key: cord-346245-o9hvuwvq authors: Harvey, David J. title: Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2009–2010 date: 2014-05-26 journal: Mass Spectrom Rev DOI: 10.1002/mas.21411 sha: doc_id: 346245 cord_uid: o9hvuwvq This review is the sixth update of the original article published in 1999 on the application of MALDI mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings coverage of the literature to the end of 2010. General aspects such as theory of the MALDI process, matrices, derivatization, MALDI imaging, arrays and fragmentation are covered in the first part of the review and applications to various structural typed constitutes the remainder. The main groups of compound that are discussed in this section are oligo and polysaccharides, glycoproteins, glycolipids, glycosides and biopharmaceuticals. Many of these applications are presented in tabular form. Also discussed are medical and industrial applications of the technique, studies of enzyme reactions and applications to chemical synthesis. © 2014 Wiley Periodicals, Inc. Mass Spec Rev 34: 268–422, 2015. This review is a continuation of the six earlier ones in this series (Harvey, 1999 (Harvey, , 2006 (Harvey, , 2008 (Harvey, , 2009 (Harvey, , 2011 (Harvey, , 2012 on the application of matrix-assisted laser desorption/ionization mass spectrometry (MALDI) mass spectrometry (MS) to the analysis of carbohydrates and glycoconjugates. It is intended to bring the coverage of the literature to the end of 2010. Although the intention is to be a comprehensive as possible, there is an increasing tendency to publish MALDI data in Supplementary information. Because of the ever-increasing number of papers and journals, it has not been possible to check all supplementary information and, consequently, papers that do not refer to MALDI in the main text may well have been omitted. Also omitted are papers that simply report the mass of glycoproteins and those concerned with nucleotides and nucleosides: the latter compounds, although containing carbohydrates are considered to be different types of compound. MALDI continues to be a major technique for the analysis of carbohydrates; Figure 1 shows the year-by-year increase in papers reporting use of the technique for the period 1991-2010. Because the review is designed to complement the earlier work, structural formulae, etc. that were presented earlier are not repeated. However, a citation to the structure in the earlier works is indicated by its number with a prefix designating the review containing the structure (i.e., 1/x refers to structure x in the first review and 2/x to a structure in the second). A large number of books and review articles directly concerned with, or including MALDI analysis of carbohydrates and glycoconjugates, have been published during the review period. Those of a general nature are listed in Table 1 ; those concerned with specific carbohydrate types are listed in the appropriate sections. Details of the MALDI process are still not fully understood and several investigators have attempted to obtain greater understanding. Although not all of these studied have involved carbohydrates, they are included here because MALDI processes for different compounds are likely to be similar. One of the commonly accepted models for the formation of analyte ions in MALDI-MS assumes a primary ionization of the matrix, for example, by photoionization leading, among other things, to stable protonated and deprotonated matrix ions. Peptide and protein ions are then envisaged as being formed by secondary proton transfer reactions in the expanding matrix plume. This model has been checked experimentally by Hillenkamp et al. (2009) by comparing the yield of positive to negative ions of three peptides (bradykinin, angiotensin I and fibrinopeptide A) and six matrices a-cyano-4-hydroxycinnamic acid (CHCA, 1/23), 2,5dihydroxybenzoic acid (DHB, 1/26), 6-azo-2-thiothymine (ATT, 1/45), 4-nitroaniline (4-NA, 3/3), 2-amino-5-nitro-4-picoline (ANP, 1) and 5-aminoquinoline (5-AQ, 2), differing in gas-phase basicity by about 100 kJ/mol. It was shown that the observed ion yields cannot be explained by any single and consistent set of parameters such as gas-phase basicity or acidity of the analyte and matrix. It was concluded that the existing simple model needs be modified to fully explain the experimental findings. Liu et al. (2009a) have used synchronized dual-polarity MALDI MS to demonstrate incoherent production of positive and negative matrix ions. In both positive and negative ion modes, matrix ions were found to appear from thin, homoge-neous DHB matrix films at different threshold laser fluences. The presence of singly charged molecular matrix ions suggests that the existence of DHB ion-pairs may not be a prerequisite in the MALDI process. Photoelectrons induced by the laser excitation may assist the production of negative DHB ions, as shown in experiments conducted with stainless steel and glass substrates. At high laser fluences, the relative yield of positive and negative matrix ions remained constant when homogeneous matrix films were used, but the yield fluctuated significantly with inhomogeneous crystal morphology. This result was also inconsistent with the hypothesis that matrix ion-pairs are essential primary ions. Thus, results from both low and high laser fluences suggest that the production of positive and negative matrix ions in MALDI may occur via independent pathways. The same authors have examined the initial ionization reaction in MALDI based on the appearance of photoelectrons. The threshold laser fluence for the ejection of photoelectrons from DHB, sinapinic acid (1/48) and 2,4,6trihydroxyacetophenone (THAP, 1/44) on stainless steel targets was found to be 0.05, 0.41, and 8.39 mJ/cm 2 , respectively. These values were considerably lower than those for MALDI ions, indicating that the electron detachment probably precedes other ionization reactions. The stainless steel target was thought to General review with a section on glycolipids - Glycan analysis by mass spectrometry Short review, MALDI and ESI, applications to N-linked glycans 20 (Sekiya & Iida, 2008) Deciphering carbohydrate structures by ion mobility MS Short general review of glycomics and ion mobility. Applications to flavonoids, GAGs and glycoproteins 96 Modern MALDI-TOF mass spectrometry Development of TOF mass spectrometers since the introduction of MALDI 26 (Vestal, 2009) MALDI Mass spectrometry of carbohydrates Short general review (In Chinese) - play an insignificant role in the production of photoelectrons because suspended DHB produced a photoelectron signal similar to DHB on the surface. In addition, decreasing the DHB thickness on the target reduced the photoelectron intensity. For crystalline DHB and sinapinic acid, the photoelectron intensity was found to increase with the laser fluence (nitrogen laser at 337 nm) in less than a second order relationship, suggesting considerable reductions of ionization potentials in comparison with free molecules. According to ab initio calculations, the ionization potential of DHB clusters was found to reduce as the cluster size increased from monomer to octamer. The paper discusses the impact of these abundant electrons on ion production in MALDI. The earlier rate equation model for MALDI ion formation and reaction (Knochenmuss, 2002 (Knochenmuss, , 2003 , has been extended to include positive and negative ions of both matrix and analyte (Knochenmuss, 2009) . The resulting positive/negative ratios of secondary analyte ions show that a recent static equilibrium approach is not adequate for quantitative analysis of MALDI experiments. Although the ion ratios remain close to unity whenever the reaction free energies are at least moderately favorable, deviations from this condition result in unequal ratios of oppositely charged ions and show once again that the dynamic aspects of MALDI cannot be neglected. Molecular dynamics simulations of MALDI have been performed to investigate laser pulse width and fluence effects on primary and secondary ionization process. At the same fluence, short (35 or 350 psec) pulses were found to give much higher initial pressures and ion concentrations than longer ones (3 ns). These differences were found not to persist because the system relaxes towards local thermal equilibrium on a nanosecond timescale. Higher fluences were found to accentuate the initial disparities. Axial velocities of ions and neutrals were found to span a wide range and to be fluence-dependent. The total ion yield was found to be only weakly dependent on the pulse width and to be consistent with experimental estimates. Secondary reactions of matrix cations with analyte neutrals were efficient even though analyte ions were ablated in clusters of matrix (Knochenmuss & Zhigilei, 2010) . Lai et al. (2010) have employed transition state theory for modeling the desorption of surface ions, assuming chemical and thermal equilibrium in the solid state prior to desorption. The method was different from the use of conventional models that assume chemical equilibrium in the gas phase. This solid-state thermodynamic interpretation was used to examine the desorption of THAP and of an angiotensin I/THAP mixture. It successfully described the changes in ion yield with the effective temperature under various laser fluence and initial temperature conditions. The analysis also revealed the key role played by ion concentration in the modeling used to provide the best fit of the model to observations. Divergence of the ion beam with laser fluence was also examined using an imaging detection method and the signal saturation normally seen at high fluence was appropriately reduced by ion focusing. Simplified but deceptive theoretical interpretations were obtained when the analysis was conducted without adequate calibration of the instrument bias. The laser plume produced by several ionic liquid matrices has been studied by a post-ionization approach in which the neutrals in the ablation plume were ionized with a second laser pulse. It was found that after the initial event that produced the ions, a second, time-delayed, ablation event occurred in which the plume contained only neutral molecules. The presence of these neutral molecules was explained by a reflected-shockwave model in which the shockwave emerging from the laser ablation is reflected from the sample plate behind the sample. It was assumed that when the shockwave arrived at the sample surface it caused a second ablation of the neutral molecules (Hellwig et al., 2009) . The 355 nm multiphoton dissociation and ionization of 2,5-dihydroxyacetophenone (DHAP, 1/43) has been studied (Dyakov et al., 2009) . The experimental results indicate that photoionization that occurs in the gas phase after DHAP vaporizes from the solid phase may not play an important role in the MALDI process. A combined IR-MALDI ion source with an electrospray ionization (ESI) emitter for post desorption ionization has been described (Sampson, Murray, & Muddiman, 2009) . The source produced multiply charged ions from proteins but singly charged ions from carbohydrates (O-glycans cleaved from mucin (MUC) were tested) and avoided the fragmentation produced by some other techniques. IR-MALDI MS with a laser emission in the 6 mm wavelength range, which utilized energy absorption at the CO double-bond stretch region, has been investigated for analysis of several types of biomolecule. The softness of IR-MALDI MS was evident in the negative ion mode where abundant [MÀH] À ions were obtained for acidic biomolecules with sulfate, phosphate, or carboxylate groups. Better sensitivity was obtained than with ultraviolet (UV) MALDI MS. Furthermore, there was no substantial loss of sialic acid due to the prompt fragmentation from sialylated glycoconjugates such as gangliosides. Such loss is a common problem with MALDI analysis of sialylated carbohydrates. The technique was used in conjunction with a potent new matrix, oxamide (3), resulting in the use of low laser fluence, and removing one of the limitations of MALDI MS for biomolecular analysis of UV-MALDIsensitive molecules (Tajiri, Takeuchi, & Wada, 2009a) . Tu and Gross (2009) have reviewed methods for miniaturizing sample spots for MALDI analysis. Topics include minimizing sample dispersion by target modification, the use of hydrophobic materials as MALDI-plate surfaces, the use of microdispensing devices such as piezoelectric dispensers and the use of droplet charging by induction or polarization. A simple device for MALDI sample preparation, based on the spraying of matrix/sample solution through a stainless steel sieve, has been used for the preparation of MALDI samples of peptides, polysaccharides (PSs) and high molecular weight (MW) proteins (Cristoni et al., 2009) . The spectra obtained by laser irradiation of the resulting microspots exhibit resolution and sensitivity higher than those achieved by the commonly employed dried droplets method. Furthermore, the target surface was more homogeneous than those obtained by the dried droplet method. CHCA and super-DHB (DHB þ 2-OH,5-OMe-benzoic acid) (s-DHB) matrices were used and applications were to insulin and dextran oligomers. Ultrasound produced by a simple piezoelectric device has been used as an alternative method for soft ionization of biomolecules. Cavitation was proposed as the major mechanism producing the ions and the technique was applied to carbohydrates, proteins and fatty acids. However, although an abundant ion, said to be the [MþH] þ from the high-mannose glycan Man 8 GlcNAc 2 (5/20), was obtained in the presence of DHB, THAP, or sinapinic acid, neither the mass nor the stated elemental composition were consistent with this structure . The recent availability of commercial ion mobility instruments offers another dimension to carbohydrate analysis by providing the ability to separate by molecular shape and offering the possibility of rapid isomer differentiation. However, resolutions on current instruments are comparatively poor and do not match those that can be obtained with high performance liquid chromatography (HPLC). A comparison of three types of ion mobility MS (field asymmetric waveform ion mobility (FAIMS), drift tube and traveling wave ion mobility spectrometry (TWIMS)) for separation of chiral molecules with applications to monosaccharides has been reported (Enders & McLean, 2009) . Data are best reported as rotationally averaged collisional cross-sections. These can be obtained directly with drift tubes and indirectly with the TWIMS instruments. Obtaining such measurements with FAIMS instruments, however, is more challenging. Collisional-cross sections have been measured for a large number of biologically relevant molecules including oligonucleotides (n ¼ 96), carbohydrates (n ¼ 192), lipids (n ¼ 53), and peptides (n ¼ 610). Collisional cross sections increased with mass but were found to be different for each molecular type in the order oligonucleotides < carbohydrates < peptides < lipids. The specific correlations were best described by logarithmic regressions. Thus, the technique was able to separate compounds of different structures but with the same or similar MWs. In addition, some separation of compounds with the same mass within a particular class was possible. The latter point was demonstrated by separations of isobaric oligonucleotides, which were interpreted by molecular dynamics simulations McLean, 2009) . have used ion mobility to extract carbohydrate ions from incubation mixtures obtained from protein-N-glycosidase (PNGase) F release of N-glycans. Glycoproteins such as ribonuclease B (RNaseB) were first digested with trypsin followed by PNGase F and then analyzed directly by ion mobility MS with a Waters Synapt instrument. The carbohydrate ions showed different mobilities from peptide and other contaminating ions allowing them to be extracted directly from the crude mixtures. Both ESI and MALDI ion sources were used; the MALDI ion source was better at ionizing the larger carbohydrates and did not suffer from the problem of producing both [MþH] þ and [MþNa] þ ions that were seen in the ESI source. The technique would appear to have great potential for rapid glycan analysis, particularly as the Synapt instrument also offers the ability to fragment the isolated ions. Hossain and Limbach (2010) have reviewed matrices used for MALDI analysis of several compound classed including carbohydrates. The review covers common matrices such as DHB and CHCA and less common systems such as liquid matrices and carbon nanotubes (138 references). The use of ionic liquid matrices has also been included in a review by Liu et al. (2009e) although the bulk of this review discusses the use of ionic liquids for sample preparation. The thermal stability of several commonly used crystalline matrices, 2,5-DHB, THAP, CHCA, sinapinic acid, nor-harmane (1/35) and harmane (1/34) has been studied by heating them at their melting point and characterizing the remaining material by a variety of spectroscopic and chromatographic techniques. In general, all compounds, except for CHCA and sinapinic acid, remained unchanged after fusion. CHCA showed loss of CO 2 , yielding a trans-/cis-4-hydroxyphenylacrylonitrile (4) mixture. Sinapinic acid showed mainly cis-to-trans thermal isomerization and, with very poor yield, loss of CO 2 (Tarzi, Nonami, & Erra-Balsells, 2009) . 1H-Pteridine-2,4-dione (lumazine, 5) has been described as a good matrix for phospholipids where the presence of cationcontaining compounds suppresses signals from neutral compounds. Phosphatidyinositol was reported to give a signal that was an order of magnitude higher than that obtained from DHB (Calvano, Carulli, & Palmisano, 2010) . A number of papers have reported the use of various nanoparticles as matrices. Unlike traditional chemical matrices, these materials generally produced little or no signal in the low mass region, thus making them ideal for analysis of small carbohydrates. Titanium dioxide micro-and nano-particles, prepared by hydrolysis of Ti butoxide and maintained in aqueous solution, have been evaluated as matrices for the detection of several small molecules. Most of the reported applications were to lipids but the nanoparticles were also applied to flavonols/anthocyanins and their glycosides in rose petals. The spectra showed ions from many more small molecules than did spectra recorded from DHB, particularly in the region of the DHB matrix ions. One advantage of the nanoparticles is that their small size (average of about 200 nm) facilitates their penetration into tissue for in situ imaging (Lorkiewicz & Yappert, 2009) . Titanium dioxide (TiO 2 ) has also be used by Kawasaki, Okamura, and Arakawa (2010) for ionization of several types of compound, including carbohydrates, with a similar absence of low mass matrix ions. In a study of the most suitable crystalline form, the anatase-type TiO 2 , was shown to be the best. Nanoparticles of diamond, TiO 2 , titanium silicon oxide, barium strontium titanium oxide, and silver have been examined for their potential as MALDI matrices for direct laser desorption/ionization of carbohydrates, especially fructans, from plant tissue. Two sample preparation methods including solventassisted and solvent-free (dry) deposition were compared. All examined nanoparticles except Ag were found to desorb/ionize standard sucrose (6) and fructans in both positive and in negative ion mode. In positive ion mode, sugars gave [MþNa] þ and/or [MþK] þ ions depending on the ionic composition of the sample spot, and [MÀH] À ions in negative mode. Ag nanoparticles yielded good signals only for non-salt doped samples that were measured in the negative ion mode. When used to study fructans in plant tissues all the nanoparticles except Ag could desorb/ionize carbohydrates in both ion modes. nanoparticles gave similar limit of detection (LOD) for standard fructan triose (1-kestose, 7) in the positive ion mode and better LODs in the negative ion mode than those given by the common crystalline organic MALDI matrices such as DHB, nor-harmane or carbon nanotubes. Although lower signal-to-noise ratio (S/N) signals were obtained from the tissues with solvent-free matrix deposition than when solvent was used, the reproducibility averaged over all sample was more uniform (Gholipour et al., 2010) . MALDI detection at the level of several hundred zmoles has been achieved by the addition of gold nanoparticles (AuNPs) with a diameter of several tens of nanometers into a sample solution (Shibamoto et al., 2009) . N-Acetyl-tetraose 1 fmol/mL gave a strong signal in the presence of 50 nm AuNP (4.5  10 7 particles). The mechanism appears to be related to surface plasmon (SP) excitation of the AuNPs. Citrate-capped AuNPs have been shown to act as matrices for the determination of several types of biomolecule, including carbohydrates such as starch, dextrins, and glycosphingolipids (GSLs) in the presence of high concentrations of salt (Wuhrer, Koeleman, & Deelder, 2009b) . With the GSLs, however, some loss of sialic acid (1/11) was found. A combination of immobilized silica and DHB on iron oxide magnetic nanoparticles has been shown to give a clean background and to be appropriate for the analysis of a range of small molecules, including glycolipids . The ratio between SiO 2 and DHB was found to affect the surface immobilization of DHB on the nanoparticle, critically controlling the ionization efficiency and background. Enhancements of molecular ion signal over those produced from DHB alone were noted and high quality product-ion spectra were obtained. Fullerene (8)-derivatized silica (pore size 30 nm) also appears to be good for small molecules including monosaccharides (Szabo et al., 2010b) . Multifunctional ZrO 2 nanoparticles and ZrO 2 -SiO 2 nanorods have been successfully used as matrices for cyclodextrins in atmospheric pressure and vacuum MALDI. The LOD for cyclodextrins was found to be 7.5-20 fmol and the matrices were used to analyze cyclodextrins in urine samples (Kailasa & Wu, 2010) . Manganese dioxide nanoparticles have been used to ionize ginsenosides with the advantage that the spectra lacked matrix ions in the low mass region allowing low-mass postsource decay (PSD) ions to be clearly visible. The technique was named nanoparticle-assisted laser desorption/ionization (nano-PALDI) (Sahashi, Osaka, & Taira, 2010) . Single-crystalline EuF 3 hexagonal microdisks with hollow interiors have been prepared as background free matrices. The long-lived excited state of europium ions is capable of transferring energy to the molecules allowing the microdisks to act as matrices. They were successfully used for analysis of small peptides, amino acids and hydroxypropyl b-cyclodextrin (9) (Chen et al., 2009d) . Another new matrix with low background is mesoporous silica, SBA-15, functionalized with quinoline . The material was obtained by using calcined . Compared with DHB and SBA-15 itself, the modified material demonstrated several advantages in the analysis of small molecules such as less background interference ions, high homogeneity, and better reproducibility. Detection limits in the region of 8 pmol were reported. In a more traditional approach for reducing low-mass matrix ions, Fujita et al. (2010) have used b-cyclodextrin (4/6) mixed with THAP or 2,4-DHAP. The latter compounds, in particular, formed inclusion complexes with the cyclodextrin as demonstrated by the lack of a similar effect when the corresponding linear carbohydrate, maltoheptaose (11) was used as a co-matrix. A method for recording negative ion spectra that is suitable for small molecules in that it produces no matrix-related ions in the low mass region uses the proton sponge, 1,8-bis(dimethylamino)naphthalene (DMAN, 12) as a solution in ethanol as the matrix. This compound has very high proton affinity and can take up protons from even weak acids to form deprotonated anions. Moreover, DMAN in solution has a strong UV absorption band in the region 330-350 nm, fully compliant with frequencies of nitrogen and neodymium-doped yttrium aluminium garnet (laser) (Nd:YAG) UV lasers. The matrix was found to be suitable for a range of small molecules including fatty acids, carbohydrates and prostaglandins (Shroff & Svato, 2009; Shroff & Svatos, 2009; . The authors proposed to name the technique as matrix-assisted ionization/laser desorption, abbreviated as MAILD, MS . N(CH 3 ) 2 (CH 3 ) 2 N 1,8-Bis(dimethylamino)naphthalene, 12 Zhang et al. (2010i) have reported a new method for the analysis of small molecules by using matrices such as metalphthalocyanines (MPc, 13) which form matrix-analyte adducts and shift the molecular ions into a high and interference-free mass range. The mass of the target analyte was calculated by subtracting the mass of MPc from the mass of the MPc-analyte adduct. The MPcs themselves were also detectable and could serve as internal standards. Various MPcs with aromatic or aliphatic groups and different metal centers were explored. Aluminum-phthalocyanines (AlPcs), gallium-phthalocyanines (GaPcs), and indiumphthalocyanines (InPcs) were found to be efficient matrices for a large number of compound types and formed MPc-analyte adducts in either the positive or negative ion mode. The detection limits varied from 17 to 75 fmol, depending on analyte. Mass Spectrometry Reviews DOI 10.1002/mas Graphene has also been reported to be a good matrix for low MW compounds with essentially no ions from the matrix (Dong et al., 2010) . Colloidal graphite has also been used for atmospheric pressure MALDI and has been shown capable of producing both [MþH] þ and [MÀH] À from many types of small molecules, including glycosylated flavonoids (Perdian, Schieffer, & Houk, 2010) . Two reviews on ionic liquids, including their use as MALDI matrices have been published (Soukup-Hein, Warnke, & Armstrong, 2009; Sun & Armstrong, 2010) In an extensive study to find new liquid matrices 114 matrices were tested and 105 new ionic liquids were prepared (Crank & Armstrong, 2009) . Both the anionic and cationic moieties were varied systematically to find a matrix with the best physical properties, analyte signal intensity and widest mass detection range. The developed matrices showed a wide mass detection range (<1,000 Da to >270,000 Da) for proteins and peptides with greater S/N ratios than solid matrices and could effectively ionize proteins of large mass without disrupting noncovalent interactions between monomers. It was found that both the proton affinity and pK a of the cation have a large effect on the ability of the matrices ionize the analyte. The matrices could be used to detect carbohydrates with fewer degradation products than solid matrices. N,N-diisopropylethylammonium a-cyano-4-hydroxycinnamate and N-isopropyl-Nmethyl-t-butylammonium a-cyano-4-hydroxycinnamate were the best matrices for proteins and peptides, while N,N-diisopropylethylammonium a-cyano-4-hydroxycinnamate and N,Ndiisopropylethylammonium ferulate were found to be the best matrices for carbohydrates. Another novel ionic liquid matrix has been made from the 1,1,3,3-tetramethylguanidinium (14) salt of THAP and found to be well suited for the MALDI analysis of glycopeptides and glycans, particularly as it appeared to overcome the well-known ionization suppression of carbohydrates in the presence of peptides. For example, even glycopeptides containing short peptide backbones and large carbohydrate moieties gave high signal intensities when using this matrix even though they did not produce spectra directly from THAP. In a second experiment, glycans were released with PNGase F from total tryptic digests derived from glycoproteins and analyzed by MALDIquadrupole ion trap (QIT)-time-of-flight (TOF). When using the liquid matrix, it was possible to detect the glycans with high intensities in the presence of the tryptic peptides, whereas, once again, glycan ionization was completely suppressed when measured with THAP alone. The extent of metastable decay of glycopeptides was also found to be reduced when using the liquid matrix (Ullmer & Rizzi, 2009) . Ionic liquid matrices have shown considerable advantages over conventional matrices for MALDI-MS analysis of polysulfated carbohydrates such as heparin and heparin sulfate. These compounds are not easily analyzed by UV-MALDI MS analysis because of the thermal lability of their O-and N-SO 3 moieties. Two new ionic liquid matrices based on the combination of 2-(4-hydroxyphenylazo)benzoic acid (HABA, 1/32) with 1,1,3,3-tetramethylguanidine or spermine (1/30) have been successfully applied to the analysis of these compounds (Przybylski et al., 2010b) . These matrices gave improved signalto-noise ratios as well as a decrease of fragmentation and desulfation. Sulfated oligosaccharides were detected with higher sensitivity than with the usual crystalline matrices, and their intact sulfated ions [MNa] À were easily observed. MALDI-MS characterization of challenging analytes such as heparin octasaccharide carrying eight O and four N-sulfo groups, and heparin octadecasulfated dodecasaccharide was also successfully analyzed. In a paper published in 2007, Gomenez et al. (2007) reported that DHB, with vacuum drying to improve target spot homogeneity, was a better matrix than sinapinic acid for obtaining reliable molecular mass values of intact glycoproteins because it prevented sugar fragmentation. In a follow-up to this work, the group have investigated the use of liquid matrices prepared from DHB and sinapinic acid with the aim of avoiding the vacuum drying step (Giménez et al., 2010) . The best results were obtained with a variety of glycoproteins such as bovine a1acid glycoprotein, bovine fetuin and human transferrin (TF) from matrices prepared by adding an equimolar amount of butylamine (64.1 mL) to 3,242 mL of a 200 mM solution of sinapinic acid or DHB in MeOH. The mixture was vortexed and sonicated for 1 min, evaporated to approximately 100 mL with air, and finally reconstituted with 100 mL of EtOH or MeCN, for DHB and sinapinic acid mixtures, respectively. Furthermore, it was noted that both matrices gave the same masses for the tested glycoproteins that agreed with literature values, suggesting that no fragmentation of the carbohydrate moieties had occurred. A matrix consisting of CHCA and aniline (15) has also proved to be successful for a number of different compounds, including raffinose (16) (Calvano, Carulli, & Palmisano, 2009) . Although it was noted that this matrix gave a stronger signal than CHCA alone, the latter matrix is not very efficient in ionizing carbohydrates. A comparative investigation of several matrices for analysis of the small carbohydrates, glucose (Glc, 1/4) and sucrose (6) has been performed by Yang et al. (2010c) . Of the matrices studied, sodiated DHB, carbon nanotubes, an ionic liquid matrix of DHB-pyridine, a binary matrix of DHB-aminopyrazine (6/7), zinc oxide nanoparticles and AuNPs, the best sensitivity was provided by the carbon nanotubes. The detection limit was 3 pmol. Both carbon nanotubes and the ionic liquid matrix exhibited the highest reproducibility. Tzeng, Zhu, and Chang (2009) have investigated doping various MALDI matrices with alkali metal hydroxides. It was found that for neutral underivatized oligosaccharides, the addition of 2% NaOH to DHB caused partial alkaline degradation by glycosidic cleavages upon laser desorption. The effect intensified when nonacidic compounds such as THAP or 5-amino-2-mercapto-1,3,4-thiadiazole (AMT, 4/7) were used as the matrix. The degradation allowed identification of the reducing end residue of the analyte and facilitated its structural characterization by PSD TOF-MS. Use of matrices consisting of LiOH and THAP or AMT led to the production of singly as well as multiple lithiated ions. Multiple lithiation appeared to occur with carbohydrates containing free 3-OH groups. Up to 6 Li atoms were found to be incorporated for maltoheptaose, b-cyclodextrin, and dextran 1500. Such a "lithium tagging" technique makes it possible to differentiate positional isomers of milk-neutral oligosaccharides, lacto-N-difucohexaose I and II (LNDFH-I (17) and LNDFH-II, (18)), without the need of chemical derivatization or tandem MS analysis. LNDFH-II, 18 = GlcNAc, = galactose, = fucose, = glucose (for details, see (Harvey, et al., 2009c) Choi and Lee (2009) have studied the ionization efficiencies of maltooligosaccharides (degree of polymerization, DP 1-7) with the cations Na þ , Li þ , and K þ in salts containing TFA À , Cl À , and OH À with DHB as the matrix. The signal strength rose with the number of glucose units with sodium consistently giving the most intense signals. The nature of the anion also affected the ionization with the TFA À salts generally being the most effective. A sample preparation method developed by Nishikaze and Amano (2009) has been compared with the conventional drieddroplet or ethanol (EtOH) recrystallization methods and reported to give superior results in terms of ion yield and signalto-noise ratio. The method, named the "reverse thin layer method" consists of first, complete drying of the oligosaccharide solution on the MALDI target plate and then deposition of the matrix dissolved in a small amount of MeOH. Using this method, a relatively homogeneous matrix crystal was generated and higher yields of both positive and negative ions were obtained. The authors comment that the method could be applied to various other matrices including both solid and liquid matrices. A method for direct archaebacterial glycolipid and lipid analysis by MALDI MS in intact membranes, without prior extraction/separation steps, has been developed by Angelini et al. (2010) . The purple membrane isolated from the extremely halophilic archaeon Halobacterium salinarum was used as a model, lyophilized and ground with 9-aminoacridine (9-AA, 6/ 18). The mixture was crushed in a mechanical die press to form a thin pellet, small pieces of which were attached directly to the MALDI target. In parallel, solution spectra of individual phospholipids and glycolipids, were analyzed by MALDI-TOF/ MS using 9-AA as the matrix. Results show that 9-AA is a suitable matrix for the conventional MALDI-TOF/MS analysis of lipid extracts from archaeal microorganisms, as well as for fast and reliable direct dry lipid analysis of lyophilized membranes. A new technique termed matrix-free material-enhanced laser desorption/ionization mass spectrometry (mf-MELDI-MS) has been described which employs a single compound prepared by immobilizing bradykinin on silica gel coupled to 4-(3triethoxysilylpropylureido)azobenzene (19) for both the MALDI matrix and a stationary phase for thin-layer chromatography (TLC). The technique was applied to the analysis of carbohydrate reference standards such as glucose, sucrose, raffinose and plant extracts from Quercus robur (oak). Carbohydrates formed [MþNa] þ and abundant [MþK] þ ions. The MELDI material adsorbs the laser energy sufficiently for desorption and ionization and delivered excellent results in respect to signal-to-noise (S/N) ratio (S/N ratio: >9/1) and sensitivity with a LOD in the low ng/mL range. For preparation of the TLC plates, the modified silica gel was suspended in methanol, acetone, acetonitrile or acetonitrile/water (1:1) for 3 min. About 15-20 mL of the suspension was applied in narrow channels cut into a stainless steel target in the form of a thin layer and dried at room temperature. The sample was placed on this layer for subsequent TLC in n-BuOH/acetone/acetic acid/ H 2 O (35:35:10:20) as mobile phase (Qureshi et al., 2010) MALDI imaging has seen many developments during the review period and several reviews and related articles have been published; details are in Table 2 . A mass spectrometer with ion mobility separation capability (Waters Synapt) has been used by Snel and Fuller (2010) to produce high spatial resolution images of glucosylceramide (20) in the spleens of mouse models of Gaucher disease. The matrix (CHCA) was applied to the tissue sections with an airbrush. For data acquisition, the laser was continually fired at one position until no more ions were observed and then the sample was moved by 15 mm (laser diameter about 150 mm). Ions were generated from only the un-irradiated surface at each of these positions achieving an effective spacing of 15 mm. At this spacing, it was possible to visualize macrophages enriched in glucosylceramide which could be distinguished from other cell types in the spleen. Current MALDI mass spectrometric imaging (MSI) spatial resolution is typically limited by the diameter of the laser spot-size, which is usually between 50 and 100 mm, covering an area equivalent to tens of mammalian cells. Perdian and have addressed the problem of acquiring high resolution imaging with high resolution MS on an Orbitrap mass spectrometer. At a spatial resolution of 100 mm, the Orbitrap mass analyzer is able to image a 2,000  2,000 mm 2 sample area in 7-14 min with one scan per step, depending on the resolution. If the spatial resolution is increased to 5 mm, the same size sample area will take more than 44-88 hr to complete the experiment, a time that is not practical in the normal laboratory. However, because the Orbitrap also has a linear ion trap (IT) analyzer, this was utilized, together with a two-motion plate movement to reduce the time while maintaining the resolution. Thus, for every pixel position on the target, the laser spot was moved in a spiral fashion such that both Orbitrap and MS/MS data were acquired. With a fast Nd:YAG laser the data acquisition time was decreased by 43-49% compared to that from orbitrap-only scans; however, 75% or more time could be saved for higher mass resolution and with a higher repetition rate laser. Using this approach, a high spatial resolution of 10 mm was maintained at IT imaging, while Orbitrap spectra were acquired at a lower spatial resolution, 20-40 mm, all with far less data acquisition time. Furthermore, various MS imaging methods were developed by interspersing MS/MS and MS fragmentation n times (MS n ) IT scans during Orbitrap scans to provide more analytical information. The method was applied to several flavonol glycosides from an Arabidopsis flower petal in which MS/MS, MS n , IT, and Orbitrap images were all acquired in a single data acquisition. Spectra were acquired in negative mode and no matrix was required. For UV-absorbing compounds such as flavonoids and their glycosides, a matrix is not necessary for imaging as demonstrated by imaging at the single cell level of secondary metabolites in Arabidopsis thaliana and Hypericum species (Hölscher et al., 2009) . The highest spatial resolution achieved, 10 mm, was much higher than that achieved by commonly used MALDI) imaging protocols. A new matrix-free technique called nanostructure-initiator mass spectrometry (NIMS) has been developed for the analysis e c n e r e f e R s n o i t a t i C s t n e t n o C t c e j b u S General review Short article, mainly applications 50 (Murayama et al., 2009) General review Methods and applications to peptides and lipids (including glycolipids) 96 General review Short general review (Award lecture) 47 General review From origins to state of the art 41 (Francese et al., 2009a) Concise review of mass spectrometry imaging (Sugiura et al., 2010b) and tissue imaging of carbohydrates and steroids (Patti et al., 2010) . Analysis was accomplished by spray depositing NaCl or AgNO 3 with a fused-silica PicoTip emitter onto a porous silicon surface to provide a uniform layer rich with cationization agents prior to desorption of the fluorinated polymer initiator. Upon laser irradiation, carbohydrates produced [MþNa] þ ions whereas steroids formed [MþAg] þ . For glucose, a plot of concentration against the 12 C/ 13 C 6 ratio was linear with a correlation coefficient R 2 of 0.9975 over the range 1-200 mM. Carbohydrates and steroids could be detected down to the 800-amol and 100-fmol levels, respectively. The ability of the method to perform tissue imaging was demonstrated by imaging the distribution of sucrose in a Gerbera jamesonii flower stem and the distribution of cholesterol (21) in a mouse brain. The flower stem and brain sections were placed directly on the ion-coated NIMS surface without further preparation and analyzed directly. No deposition of a matrix onto the sample surface was needed. A similar matrix-free method, termed nano-assisted laser desorption-ionization (NALDI) MS for tissue imaging has been demonstrated by Vidová et al. (2010) . Commercially available nano-structured surfaces were used as substrates for imprinting tissue sections. The lithographic transfers were washed and the lipids were imaged by laser desorption mass spectrometry (LDMS). The NALDI imaging of lipid transfers was compared with standard MALDI imaging of matrix-coated (CHCA) tissue sections and the resulting images were found to be of the same quality with no spatial information being lost due to the imprinting process. NALDI imaging was reported to be faster due to the absence of the time-consuming matrix deposition step, and the NALDI mass spectra were less complex and easier to interpret than MALDI spectra. The method was applied to the analysis of phospholipids, GSLs and glycerophospholipids in mouse kidney slices. NALDI MS was able to identify the same lipid species as MALDI and was reported to provide better distinction between kidney and adrenal gland tissues based on the lipid analysis. Miura et al. (2010a) have developed a MALDI imaging system that is claimed to be able to image from single cells in thin tissue sections. An indium tin oxide-coated glass slide marked with a 50 mm-wide mesh work to enabled matching of optical and MS images was used for mounting tissue sections. A suspension of HeLa cells in culture medium was mounted onto the slide and incubated for 6 hr at 37˚C. Cells were then washed with phosphate-buffered saline (PBS) and the appearance of single cells adhering to the glass was observed by optical imaging. The single cell-adhered glass slide was then mounted onto MALDI sample plate with a parallel experiment involving brain tissue slices and the matrix, 9-AA, was applied with an airbrush. Several metabolite peaks were detected at the position of the single cell. Adenosine triphosphate (ATP, 22; m/z 505.99, identified by comparison with a standard sample) was identified with a good signal-to-noise ratio. Peaks were also obtained from other metabolites such as fructose-1,6-bisphosphate (23) A major hurdle for imaging gangliosides in tissue using MS is that sialic acid residues can be dissociated in the ionization process. Chan et al. (2009a) have investigated the ionic liquid matrix CHCA/1-methylimidazol (6/62) (1:1 w/w) and have found that it produces excellent sensitivity for ganglioside detection without significant loss of sialic acid residues. The matrix was applied to the tissue samples with an airbrush; the method best adapted to handling a mixed matrix whose components have different volatilities. Image analyses of mouse brain tissue sections demonstrated that the N-fatty acyl chains of gangliosides were differentially distributed in mouse hippocampal regions. Gangliosides with N-C18 acyl chains were enriched in the CA1 region, while gangliosides with N-C20 acyl chain were enriched in dentate gyrus. Colsch and Woods (2010) have developed a method for imaging sialylated GSLs in mouse brain. The total glycolipid profile was obtained by MALDI-TOF following solvent extraction and then individual species were mapped from frozen brain slices by MALDI using a linear TOF/TOF system in negative ion mode. The matrix, which consisted of saturated 2,6dihydroxyacetophenone (DHAP, 1/49) dissolved in 50% ethanol with the addition of ammonium sulfate (125 mM) and 0.05% of heptafluorobutyric acid (HFBA) was applied with a CHIP-1000 chemical inkjet printer with a piezoelectric head. Twenty-eight nanoliters of matrix were deposited per spot with the distance between two spots of 240 mm. The ammonium sulfate in the matrix mixture limited the formation of salt adducts, while the addition of HFBA increased the stability of DHAP in the vacuum and reduced its rapid sublimation. Some sialic acid loss was noted, particularly with GD1 and GT1, which were detected as the ganglioside ( The results showed differences in GSL localization in several brain regions depending on the sialic acids and the ceramide (Cer). Imaging of lipids, including GSLs, has been reported by Landgraf et al. (2009) using a hybrid linear IT/Orbitrap mass spectrometer that allowed the acquisition of MS/MS spectra. A dramatic improvement in mass spectral resolution and a decrease in background were observed from lipids distributed within nerve tissue when compared with results obtained from fragmentation within the linear IT. The DHB matrix was applied to the dried tissue samples with an Epson inkjet printer and the MALDI ion source was operated at a pressure of about 75 mTorr. The technique was used to image lipids from rat spinal cord sections. Goto-Inoue et al. (2009) have imaged the glycolipid, seminolipid (24) in mouse testis. This sulfated glycolipid comprises more than 90% of the glycolipid in mammalian testis and disruption of the gene catalyzing transfer of galactose (Gal) results in male infertility due to the arrest of spermatogenesis. Different molecular species are defined by fatty acid composition. Tissue imaging was performed from thaw-mounted tissue slices that had been sprayed with DHB with an airbrush. It was found that the major molecule (C16:0-alkyl-C16:0-acyl) was expressed throughout the tubules: some (C16:0-alkyl-C14:0acyl and C14:0-alkyl-C16:0-acyl) were predominantly expressed in spermatocytes and the other (C17:0-alkyl-C16:0acyl) was specifically expressed in spermatids and spermatozoa. This is the first report to show the cell-specific localization of each molecular species of seminolipid during testicular maturation. Experimental details for performing imaging of glycolipids using the airbrush application method for applying DHB, as used by this group, has been published separately (Goto-Inoue, Hayasaka, & Setou, 2010a) . The distribution and localization of GSLs present in mouse brain sections have also been investigated using nanoparticleassisted laser desorption/ionization imaging MS. AuNPs modified with alkylamine were used as a new matrix to maximize the detection of GSLs. The mouse brain was frozen in liquid nitrogen, and serial sections were thaw-mounted onto indiumtin oxide (ITO)-coated glass slides. A thin layer of AuNPs or DHB matrix was applied to the surface with an airbrush. IMS analyses were performed by raster-scanning in the x-axis with a scan pitch of 200 mm in the y-axis. Sulfatides and gangliosides were detected in mouse brain sections with the new matrix whereas it was difficult to detect them using DHB (Goto-Inoue et al., 2010c ). An oscillating capillary nebulizer (OCN) was also used by Chen et al. (2010g) for analysis of sphingolipids in tissue slices in Tay-Sachs/Sandhoff disease. In addition to the above-mentioned matrix-free methods, Jung et al. (2010) have reported imaging of cellulose in poplar (Populus deltoids) stem using more traditional techniques. Analysis of microcrystalline cellulose was performed first to determine the best matrix. DHB gave much stronger signals than matrices such as CHCA or sinapinic acid and was applied to poplar cellulose with an OCN. Ions at m/z 1, 500, 2, 472, and 3, 120 (DP 9, 15, and 19) were monitored with a Voyager DE STR instrument and produced images that closely resembled the optical image. Taira et al. (2010a) , on the other hand used an airbrush with CHCA to image ginsenosides in cross-sections of Panax ginseng root and used MS/MS to obtain detailed structural information. Ginsenosides were found to be located more in the cortex and periderm than in the medulla and that they were at greater concentration in the root tip than in the center of the root. Several carbohydrates including hexoses (Hex) and D-fructose 6-phosphate have been imaged in eggplant (Solanum melongena, also known as aubergine) fruits from DHB although the paper was mainly concerned with imaging of g-aminobutyric acid (GABA, 25) (Goto-Inoue, Setou, & Zaima, 2010b) . Li, Bohn, and Sweedler (2010h) have compared two MS imaging methods, MALDI and SIMS, for glycan detection in the stems of the perennial grass Miscanthus  giganteus. Several methods of sample preparation were investigated for MALDI. A thin (2 nm) gold coating provided high quality signals of oligosaccharide-related ions and DHB also showed high efficiency for ion production. On the other hand, CHCA produced only very weak spectra, consistent with its use as a stand-alone matrix for carbohydrates. Direct laser ablation of untreated sections gave high resolution images, although coating the sections with a nanometer thick layer of gold greatly enhanced the quality of the SIMS images. Two reviews describing derivatization reactions have been published (Busch, 2010; Ruhaak et al., 2010b) , the second (207 references) is more comprehensive and deals specifically with N-linked carbohydrates. These derivatives are most often used for introducing fluorescent tags for chromatographic detection but also find use in MS. 2-Aminobenzamide (2-AB, 1/56) and 2-aminopyridine (2-AP, 1/52), favored by Japanese investigators, are the derivatives most frequently encountered; use of the latter derivatives has been reviewed by Hase (2010) . Several new derivatives (or labels) have been reported. Thus, 5-amino-2-naphthalenesulfonic acid (ANSA, 26) has been used to derivatize N-glycans by reductive amination for capillary electrophoresis (CE), HPLC, and MALDI-TOF analysis (Briggs et al., 2009) . The derivative was reported to give superior resolution in both CE and HPLC analysis to the wellused 8-aminopyrene-1,3,6-trisulphonic acid (APTS) derivatives and in MALDI-TOF analysis, the negative charge enabled both neutral and acidic glycans to be examined simultaneously. 3-Amino-9-ethylcarbazole (6/19) (Mou et al., 2009) , another new derivatizing agent, has been reported to increase sensitivity of MS detection. 2-Picoline-borane (27) has been proposed as an alternative to toxic sodium cyanoborohydride for the reductive amination reaction (Ruhaak et al., 2010a) . Pabst et al. (2009) have compared the preparation and performance of 15 different labels for N-glycan analysis. Several cleanup procedures were developed for cleaning these derivatives before analysis, of which hydrophilic interaction solidphase extraction (SPE) appeared to be the most widely used. However, the cleanup was laborious and a better method was sought. Simple addition of acetone resulted in the formation of a precipitate, which turned out to be the labeled oligosaccharide. A single addition and removal of acetone reduced the amount of reagent to approximately 0.2% (measured with 2-AB). Two further extractions of the pellet with acetone reduced the excess of amine reagent by at least as much as clean-up with a cyano-SPE cartridge. In addition, reduction of the Schiff base of 2-APlabeled glycans proceeded faster and/or required less reagent when the samples were pre-purified before the addition of reducing agent. Acetone extraction was successfully applied to many other labels such as 2-AB and 3-aminobenzoic acid (2-AA) (1/57). With respect to the performance of the individual labels, procainamide (28) emerged as more sensitive than 2-AA for normal-phase HPLC, but its chromatographic performance was not convincing. 2-AP gave the lowest retention on reversedphase and graphitic carbon columns and, thus, appeared to be most suitable for glycan fractionation by multidimensional HPLC. Most glycan derivatives performed better than native carbohydrates in MALDI and ESI MS, but the sensitivity gain was small and hardly sufficient to compensate for sample loss during preparation. Amano et al. (2009a) have labeled oligosaccharides with a pyrene derivative in order to acquire negative ion MALDI-QIT-TOFMS n spectra. The oligosaccharides were reacted with pyrene butanoic acid hydrazine (6/21) and then reduced with NaBH 4 and cleaned with a small C 18 column. The derivatives gave intense and stable molecular ions in both positive and negative ([MÀH] À ions) modes and as little as 10 fmol of pyrene-labeled oligosaccharides, such as monofucosyllacto-Nhexaose (29) gave sufficient signals for analysis. Although some fucose loss by in-and post-source occurred in positive ion mode, this loss was significantly less in negative mode. A method, termed glycan reductive isotope labeling (GRIL) has been introduced for glycan quantitation. Free glycans or those released from glycoproteins, were derivatized by reductive amination with either [ 12 C 6 ]aniline or [ 13 C 6 ] aniline. These dual-labeled aniline-tagged glycans could be recovered by reversed-phase chromatography and could be quantified by UV absorbance or MS. One labeled variety was used as the reference standard against which the test glycan, labeled with the other isotope was measured. Unlike previously reported isotopically coded reagents for glycans, the derivatives did not contain deuterium, which can sometimes be chromatographically resolved. The technique allowed linear relative quantitation of glycans over a 10-fold concentration range and could accurately quantify sub-picomole levels of released glycans (Xia et al., 2009) . On-target derivatization with the matrix 3-aminoquinoline (1/24) has yielded Schiff bases which could be measured directly in positive and negative ion mode from one single spot. The optimal conditions were 20 mg/mL of 3-AQ in a MeCNwater mixture (1:2 v/v) with 0.07% of an inorganic acid to give a pH of 5. In negative ion mode, spectra from chloride adducts of the derivatives were acquired from 1 fmol of oligosaccharide. Furthermore, PSD fragmentation in positive and negative ion mode was enhanced, providing information on oligosaccharide sequence, linkage, and branching. The method was applied to trifucosyllacto-N-hexaose (30) and trifucosyl-para-lacto-N-hexaose (31), two isomers occurring in human breast milk samples, which were easily identified and distinguished (Rohmer et al., 2010) . TFpLNH, 31 (Symbols as defined for structures 17 and 18. Anomericity not specified) Reductive amination derivatization has also been exploited in other areas, combined with their use in MS. Binding of sugar chains to proteins, viruses and cells is conveniently monitored by the surface plasmon resonance (SPR). Key to this method is the use of linker compounds for immobilization of the sugar chains to the gold-coated SPR chip. Sato et al. (2009a) have developed a novel linker molecule, named "f-mono," which combines a linker with a fluorescent moiety to allow high sensitivity monitoring of the glycans. Since the molecule (32) contains a 2,5-diaminopyridyl group and a thioctic acid group, conjugation with sugar chains can be achieved by the reductive amination reaction. The mass spectra of thee compounds contained a peak 2 Da higher than the molecular ion due to the reduction of the thioctic acid moiety providing a convenient method for identifying the glycans even using unfractionated crude samples. Immobilization of the derivatives onto gold-coated chips, and their subsequent SPR analyses were successively accomplished. Use of hydrazone formation removes the need for a reductive step to stabilize the derivative as required by Schiff base formation from primary amines. Experimental details for synthesis of the phenylhydrazone derivatives discussed in earlier reviews (Lattova & Perreault, 2003a,b; Lattova, Perreault, & Krokhin, 2004) have recently been reported in an edition of Methods in Molecular Biology devoted to glycomics . Small oligosaccharides and N-glycans from chicken ovalbumin have been converted into their biotin derivatives by incubating them with biotinamidocaproyl hydrazide (BACH, 33) (Kapková, 2009) . The derivatives imparted improved mass spectral sensitivity over that of the free glycans and, because the labeling reagent contained a biotin handle, it allowed the interaction between lectins and biotin-derivatized oligosaccharides to be investigated. Fragmentation of the N-linked glycans was dominated by Y and B/Y-type glycosidic ions. Han et al. (2010a) have used a new substituted hydrazine, 1-(4-cyanophenyl)-4-piperidinecarbohydrazide (34) and produced a derivative that increased detection sensitivity by about 10-fold over that from the underivatized glycan. The observation of [MþNa] þ ions rather than the expected [MþH] þ species suggested that the sensitivity increase was the result of increased hydrophobicity. MALDI analysis employed DHB and super-DHB; recrystallization of the super-DHB sample with acetonitrile substantially improved the signal-to-noise ratio and reproducibility. Girard's T reagent (1/55), with its constitutive cationic charge, has been used in quantitative measurements, as described below and for measuring a-galactose-containing N-glycans in porcine pig corneal endothelial cells and keratocytes (Kim et al., 2009c) . Because of the in-built charge, signal strengths from glycans of different structures were thought to be equal. This is not necessarily the case for formation of [MþNa] þ ions. Rather than reacting the reducing-terminal aldehydes of carbohydrates with amines or hydrazines, the reverse reaction can be used if the glycan is first converted into its glycosylamine (35). In fact, this type of reaction can be used directly on PNGase F-released N-glycans because these are released in this form. Chemical formation of glycosylamines generally utilizes the Kochetkov reaction in which the glycan is treated with an excess of ammonium carbonate. Unfortunately, this reaction is slow and can take up to five days for completion. To overcome this problem, Liu, Salas-Solano, and Gennaro (2009h) have used microwave assistance to speed the reaction up to as little as 90 min. Following amidation the glycans were reacted with tris-(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (36) to introduce a permanent charge on the glycan and the investigators were able to detect derivatized maltoheptaose at 2 fmol/mL by MALDI-TOF MS using DHB and CHCA matrices. Glycans from RNaseB, chicken ovalbumin and asialofetuin were also detected at high sensitivity. A potential problem arises from possible cleavage of the reducingterminal N-acetylglucosamine (GlcNAc) residue as such a reaction has recently been reported as a by-product of the Kochetkov reaction when, for example, ammonium bicarbonate at 37˚C was used (Murase & Kajihara, 2010 Several carbohydrates, including maltoheptose, blood type B antigen, pullulan and the glucan of Ganoderma lucidum have been converted into their naphthimidazole (NAIM) derivatives (37) in high yields by the iodine-promoted oxidative condensation (Scheme 1; Lin et al., 2010b) . The reaction took about 6 hr to go to completion giving derivatized carbohydrates that gave enhanced MALDI signals ([MþH] þ ions) compared with those from the free carbohydrates or their 2-AB derivatives with DHB or THAP as matrices. Less than 1 pmol of carbohydrate could be detected. Furthermore, the derivatives could easily be hydrolyzed to the parent glycans. A further series of such derivatives has been synthesized by condensation with diamines such as substituted benzene-1,2-diamine (38) in order to increase hydrophobicity and detection sensitivity (Lin et al., 2010c) . Using maltotriose (11, n ¼ 1), derivatives with pyrimidine-4,5diamine (39), pyridine-3,4-diamine (40) and 1,2,5-oxadiazole-3,4-diamine (41) B. Derivatives of Other Sites Permethylation has long been used in carbohydrate analysis, most frequently for linkage determination by gas chromatography/mass spectrometry (GC/MS) following hydrolysis and acetylation (permethylated alditols acetates). However, there appears to be an increasing trend to employ the reaction for MALDI analysis. The advantage of permethylation is that it increases sensitivity and several investigators employing its use appear to detect larger glycans, particularly N-glycans, than by the use of underivatized glycans. However, sample clean-up of the highly basic reaction mixtures can be a problem with, in some cases, losses offsetting any gain in sensitivity. Introduction of solid-phase permethylation has improved the situation. Experimental details of the solid phase permethylation method (methyl iodide on sodium hydroxide beads) (Kang, Mechref, & Novotny, 2008; Kang et al., 2005) that is capable of permethylating very small amounts of carbohydrate have been published in Methods in Molecular Biology (Mechref, Kang, & Novotny, 2009a) . On the negative side, it has been reported that permethylation can lead to loss of O-linked acetyl groups from certain sialic acids (Liu & Afonso, 2010 ). An extension of the solid-phase method to allow sulfated glycans to be analyzed by MALDI-TOF MS has been developed (Lei, Mechref, & Novotny, 2009) . Sulfated glycan samples were permethylated followed by methanolytic cleavage of the sulfate groups. The desulfated, permethylated glycans were then subjected to another permethylation step using deuteromethyl iodide to label the hydroxyl groups that were exposed by removal of the sulfates. The number of attached sulfate groups could be calculated from the mass-shift caused by the presence of the deuterium label and the position of the sulfate substitution could be determined by collision-induced dissociation (decomposition) (CID). The method was validated with linear standard glycans and used to identify sulfated N-glycans released from bovine thyroid-stimulating hormone (bTSH). Yu et al. (2009c) have shown that it is possible to permethylate sulfated glycans with methyl iodide and sodium hydroxide (Ciucanu and Kerek method) (Ciucanu & Kerek, 1984) , without loss of the sulfated glycans. The trick is to avoid the normal chloroform/water partition stage, which results in much of the sulfated material (unmethylated) partitioning into the aqueous phase. Instead, clean-up employed C18 reversedphase SPE cartridges and microtips self-packed with NH 2 -beads. The methylation reaction was capable of methylating phosphates but not sulfates, allowing them to be readily identified. Formation of methyl esters by reaction of the sodium salt with methyl iodide has frequently been used to stabilize sialic acids to MALDI analysis by eliminating the labile proton on the acid group. An alternative procedure for methyl ester formation that provides information on the sialic acid linkage directly from the MALDI spectrum has been published by Wheeler, Domann, and Harvey (2009) (Fig. 2) . The sugars were desalted, dried, dissolved in methanol and treated with 4-(4,6-dimethoxy-1,3,5triazin-2-yl)-4-ethylmorpholinium chloride (DMT-MM, 5/12). After removal of the solvent, the products were transferred directly to the MALDI target and examined from DHB. However, for the analysis of small amounts of N-glycans derived from biological sources, the method benefited from an additional clean-up stage involving the use of a Nafion 117 membrane. Unlike the reaction with methyl iodide with the sodium salt that produced a single peak from each sialylated glycan, irrespective of the linkage, the reaction with DMT-MM produced different derivatives from a2 ! 3-and a2 ! 6-linked sialic acids. The a2 ! 6-linked sialic acids produced only methyl esters whereas a2 ! 3-linked sialic acids were converted into their lactones providing a 32 Da difference in mass, thus enabling the linkage to be determined directly from the MALDI-TOF spectrum (Fig. 2) . Negative ion CID mass spectra of these neutralized glycans provided information, in many cases, on the antenna of N-linked glycans to which the variously linked sialic acids were attached. In an application of the method to the glycoprotein carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), it was shown that both a-2 ! 3-and a-2 ! 6linked sialic acids were present (Harvey, Baruah, & Scanlan, 2009a) . 4-(4,6-Dimethoxy-1,2,3-triazil-2-yl)-4-methylmorpholinium chloride in the presence of ammonium chloride, converts sialylated glycans into amides with the same linkage-specific reactivity difference. Thus, the a2 ! 3-linked sialylated glycans yield lactones, as above, whereas the a2 ! 6-linked compounds form amides. have used this reaction to examine blood serum glycoproteins but their technique differed from the above methyl ester formation in that the glycans were permethylated after reaction with DMT-MM. This reaction formed the methyl ester from the a2 ! 3-linked compounds as the result of the opening of the lactone ring, whereas the amide that was derived from the a2 ! 6-linked compounds was converted into the corresponding dimethylamide with a 13 Da increase in mass over that of the methyl ester. 4-(4,6-Dimethoxy-1,2,3-triazil-2-yl)-4-methylmorpholinium chloride can also be used to form substituted amides directly and has been used by Endo et al. (2009) to link the fluorescent derivative 2-(2-pyridylamino)ethylamine (PAEA, 42) to the carboxy group of sialic acids in sialo-oligosaccharides and gangliosides. The derivative gave good HPLC and TLC sensitivity and possessed the following advantages for MALDI analysis: suppression of preferential cleavage of Neu5Ac; enhancement of molecular-related ion intensities; simplification of MS spectra; and finally, since PAEA-amidation did not cleave the linkage between sugar and aglycon, allowed MALDI-TOF-MS and MS/ MS analyses to reveal the complete structure of the molecule. 4-(4,6-Dimethoxy-1,2,3-triazil-2-yl)-4-methylmorpholinium chloride has also been used to form amides from hexuronic acids (HexAs) in an investigation of the N-linked glycosylation of structural subunit RvH2 of Rapana venosa hemocyanin . As well as containing the rather unsusal (for N-glycans) hexuronic acid, the glycans also contained naturally methylated hexoses and internal fucose residues. Gil et al. (2010) have stabilized sialic acids by formation of amides with acetohydrazide under mild acidic conditions in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC, 43) . Glycoproteins were first reduced and alkylated and then the sialic acids were amidated. Glycans were released with PNGase F and a permanent charge was attached to the reducing terminus by further reaction with Girard's T reagent. Alternatively, derivatization with 2-AA was used and the products were examined both by HPLC and MALDI-TOF MS. The amidation reaction was performed on the glycoprotein because acetohydrazide would also have reacted with the aldehyde function of the released glycan, precluding derivatization with an amine more suited to the detection method used for monitoring the glycans. The method was applied to the analysis of N-glycans from transgenic pig-derived human factor IX. Another method for synthesis of methylamides has been reported by Liu et al. (2010i) . Sialylated glycans were reacted with methylamine in the presence of (7-azabenzotriazol-1yloxy)trispyrrolidinophosphonium hexafluorophosphate (PyAOP, 44) and N-methylmorpholine (45) for 30 min at room temperature. After methylamidation, sialylated glycans were analyzed by MALDI-TOF and ESI MS without loss of the sialic acid moiety. Both a2 ! 3-and a2 ! 6-linked sialic acids were quantitatively converted to their methylamides. This method was validated with N-glycans released from the well-characterized glycoproteins, fetuin, human a1-acid glycoprotein, and bovine a1-acid glycoprotein and was used to study N-glycans from serum glycoproteins from human, mouse, and rat. Both Neu5Ac and Oacetylated analogues were stable under MALDI conditions. Glycopeptides tend to produce weaker signals than nonglycosylated peptides and it is frequently difficult to observe their molecular ions in samples rich in unglycosylated peptides. have developed a highly sensitive on-plate pyrene derivatization method using 1-pyrenyldiazomethane (46) for acquisition of MALDI MS n spectra of glycopeptides in amounts of less than 100 fmol. Unusually, the pyrene groups were easily released from glycopeptides during ionization when DHB was used as a matrix. Thus, most ions were observed in their underivatized form. At the same time, pyrene derivatization was found to reduce the ionization of peptides and to produce signals from the glycopeptides that were strong enough for acquisition of MS n spectra that contained ions from both glycan and peptide. When the liquid matrix, 3AQ-CHCA, was used, the sialic acid linkages of the pyrene sialylated glycopeptides were found to be stable because of inefficient release of the pyrene group allowing MS n spectra of the intact glycans to be obtained. The method was used to examine glycopeptides from 1 ng of prostate specific antigen. Ohara et al. (2009) have developed a method for analysis of sulfated carbohydrates by forming complexes with 1-(pyren-1ylmethyl)guanidine (PMG, 47) from a matrix consisting of this compound and p-nitroaniline (3/3). Two types of sulfated carbohydrate were examined, chondroitin sulfate (48) and carrageenan (49). The PMG complexed with the sulfate group and was eliminated under MALDI conditions such that the molecules produced a ladder of peaks separated by masses corresponding to losses of each complexed sulfate (mass difference 353 U). In positive ion mode, the molecular ions from the chondroitin sulfates contained one more PMG molecule than the number of sulfate groups. One PMG group was presumed to bind to a carboxylate group. In negative mode, one less PMG molecule was bound. Carrageenans showed a slightly different pattern in that the number of PMG molecules found in the positive ion spectra equaled the number of sulfates. Sialic acids are classically analyzed by HPLC after formation of fluorescent 1,2-diamino-4,5-methylenedioxybenzene (DMB, 50) derivatives. These derivatives require an a-keto acid group in the sialic acid. Galuska et al. (2010b) have developed a method for specifically measuring nucleotide-activated sialic acids in the presence of unactivated acids by first reducing the keto group that is present only in the non-activated acids. Under the conditions of the derivatization reaction, the nucleotide group was removed leaving, in the case of the activated acids, only, an intact a-keto acid group that reacted with the DMB reagent. Subsequent analysis was by HPLC and MALDI-TOF. MALDI MS is used extensively in analyses with glycan arrays as summarized in recent reviews (see Table 3 ). Table 4 lists glycans that have been used in array construction. Most of the above work has been with carbohydrate preparations prior to array construction. However, MALDI MS has also been used to interrogate arrays directly. As an example, a new type of glycan array covalently or non-covalently attached to aluminium oxide-coated glass slides has been developed for studies of enzymatic reactions and protein binding . The array was prepared by tagging glycans with a polyfluorinated hydrocarbon (51) tail and spotting them robotically onto the aluminium oxide-coated slide surface which contained a layer of polyfluorinated hydrocarbon terminated with phosphonate. After incubation and washing, the noncovalent array was characterized by MALDI-TOF at a low laser energy without addition of matrix. A cellotetraose (D-Glc-(b-(1 ! 4)-D-Glc) 2 -b-(1 ! 4)-D-Glc) array was developed to study the activity and specificity of different cellulases and to differentiate the exo-and endoglucanase activities. Compared to the preparation of glycan arrays on glass slides and other surfaces, this method using phosphonic acid reacting with aluminium oxide-coated was said to be more direct, convenient and effective and represented a new platform for the highthroughput analysis of protein-glycan interactions. Polyfluorinated hydrocarbon tag, 51 In another application, a ligand affinity capture (LAC) method for detection of biotinylated biomolecules has been developed by Jørgensen, Juul-Madsen, and Stagsted (2009) . Metal-coated glass slides were treated with amino-silane and derivatized with biotin followed by avidin. Biotinylated biomolecules could then be captured and detected in the low femtomole to low picomole range by MALDI-TOF MS using CHCA in a dried droplet method. The technique was used to detect biotinylated lipopolysaccharide (LPS) and its binding to the antagonist polymyxin B. The a-mannose-specific lectin Concanavalin A (Con A) has been bound to polydopamine-modified gold, indium, and iridium surfaces and its activity was demonstrated with RNase B using SPR spectroscopy. Surface-MALDI-TOF MS experiments revealed that the affinity of the immobilized Con A depended on the oligosaccharide structure and composition. Thus Con A was found to bind certain Man 7 -(4/20), Man 8 -(5/20), and Man 9 -GlcNAc 2 RNase B glycoforms more strongly than Man 5 -and Man 6 -GlcNAc 2 glycoforms (Morris, Peterson, & Tarlov, 2009) . Concern has frequently been expressed about the ability of MALDI-TOF MS to provide quantitative information. Fortunately, this concern appears to be unjustified as two recent studies have shown. Thus, a systematic study of widely different glycopeptides was performed by Thaysen-Andersen, Mysling, and Højrup (2009) to determine the relationship between the relative abundances of the individual glycoforms and the MALDI-TOF MS signal strength. Glycopeptides derived from glycoproteins containing neutral glycans (RNaseB, IgG, and ovalbumin) were profiled and yielded excellent and reproducible quantitation (correlation coefficient r ¼ 0.9958, n ¼ 5) when evaluated against a normal-phase HPLC 2-AB glycan profile. Similarly, precise quantitation was observed for various forms of N-glycans (free, permethylated, and fluorescence-labeled) using MS. In addition, three different sialo-glycopeptides from fetuin were site-specifically profiled, and good correlation between peak intensities and relative abundances was found with only a minor loss of sialic acids (r ¼ 0.9664, n ¼ 5). For glycopeptide purification, a range of hydrophilic and graphite materials packed in microcolumn format was evaluated and proved capable of desalting without loss of quantitative information. Thus, MALDI-TOF MS signal strength of glycopeptides has been found to accurately reflect the relative quantities of glycoforms, enabling rapid and sensitive site-specific glycoprofiling of N-glycan populations. The second study relates to the concern that has often been expressed over possible losses of fucose residues when glycans are ionized by MALDI. Tajiri, Kadoya, and Wada (2009b) have recently assessed possible fucose loss and found it to be insignificant. Fucose (Fuc) is known, however, to migrate within [MþH] þ ions particularly when these are derivatized by reductive amination. Experiments on fucose loss were conducted with fucosylated glycopeptides from TF and haptoglobin. Studies with increasing collision energy on the [MþH] þ ions showed that the major fragmentation was cleavage at GlcNAc residues. Biantennary glycans containing a1,3/4-antenna fucose or a1,6-core fucose showed different fragmentation behaviors in experiments. Stability was dependent on peptide backbone sequences. Cleavage of the GlcNAcb1 ! 2Man linkage occurred at a slightly lower activation energy than for the core fucosylated (CF) species, while the linkage of a1,6core fucose was more stable than that of antenna a1,3/4 fucose. However, these fragmentations only occurred at relatively high collision energy. Consequently, the authors concluded that quantitation of fucosylated glycans by MS of glycopeptides, without collisional activation, was justified. The fucosylation levels calculated from the signal intensities in nanospray ionization and UV MALDI mass spectra were essentially the same. The mass spectrometric profiling of glycopeptides from TF from patients with congenital disorders of glycosylation (CDG-Ia and CDG-IIc) demonstrated that the elevation or reduction of fucosylation in pathological conditions can be reliably determined by MS of glycopeptides. In spite of these reassurancies, it is possible that for mixtures of compounds, complex suppression effects may degrade quantitative results. Consequently some investigators prefer LC/MS methods because the LC step removes, or minimizes, the effect that other compounds in a mixture have on the ionization of the target compounds. Instability of sialylated glycans under MALDI conditions complicates quantification and errors can possibly also be introduced by unequal ionization of glycans in mixtures. To overcome these problems with N-glycans from a therapeutic glycoprotein from a Chinese hamster ovary (CHO) cell line, Jang et al. (2009a) first formed methyl esters of the sialic acids to stabilize them and then converted the glycans to their Girard's T derivatives. These derivatives have a constitutive positive charge, thus overcoming the problem of unequal ionization. Percentages of sialylated glycans were measured at 22.5 and 5.2% in two cell lines. The results were comparable to those obtained by NP-HPLC combined with fluorescence detection using 2-AB or 2-AP derivatization. Girard's T derivatives have also been used by Kim et al. (2009a) to quantify glycans released from GSLs originating from miniature pig endothelia and islet cells. A method using a deoxyribonucleic acid (DNA) sequencer has been described for the quantitative analysis of plant Nglycans released with PNGase A or F and derivatized with APTS (1/59). MALDI-TOF analysis was used to confirm structures with the aid of exoglycosidase digestions (Lee et al., 2009i) . A method for the quantification of fructo-oligosaccharides using MALDI TOF MS with DHB as the matrix, has been developed with the fructan, raftilose, a partially hydrolyzed inulin with a degree of polymeration 2-7 as the test compound (Onofrejová, Farková, & Preisler, 2009) . Nystose (2/11), which is chemically identical to the raftilose tetramer, was used as the internal standard. Two mathematical approaches, conventional calculations and artificial neural networks (ANN), were compared for data processing. The conventional method relied on the assumption that a constant oligomer dispersion profile will change after the addition of the internal standard. On the other hand, ANN was found to compensate for a non-linear MALDI response and variations in the oligomer dispersion profile with raftilose concentration. As a result, the application of ANN led to lower quantification errors and excellent day-to-day repeatability compared to the conventional data analysis. This reproducibility was satisfactory for MS quantification of raftilose in the range of 10-750 pg with errors below 7%. The method was applied to measurements of the content of raftilose in dietary cream and it was stressed that no special optimization of the MALDI process was carried out. MALDI-TOF MS with DHB, THAP or p-nitroaniline (3/3) has been used to determine the concentrations of the unsaturated disaccharide from chondroitin sulfate (48) obtained by enzymatic digestion of native chondroitin sulfate with chondroitin ABC lyase. The signal-to-noise (S/N) ratio in the spectrum was used as a quantitative measure: amounts of chondroitin sulfate (measured as the disaccharide) down to at least 500 fmol could be detected and there was a direct correlation between the S/N ratio and the amount of chondroitin sulfate between about 2 and 200 pmol although the curve was sigmoidal. The influence of different parameters such as the matrix, the applied laser intensity and different methods of data analysis were also tested. Advantages and drawbacks of this approach are critically (Song et al., 2009d) Bacillus anthracis tetrasaccharide with thiol linker MALDI For attachment to a maleimide functionalized Microarray to study of carbohydrate-antibody interactions (Oberli et al., 2010) Glycodendrimers with N 3 group terminating in α-Man, β-GlcNAc or β-Gal TOF Immobilized on an acetylenyl-terminated gold substrate via click chemistry High-mannose glycans -Oxime linked TOF Used to probe binding to malectin MUC1 Glycopeptides TOF Synthesis on an amine-reactive hydrogelcoated microarray glass surface. To detect autoantibodies in breast cancer Correction: (Blixt et al., 2011) N-Glycan-Asn-fmoc conjugates from chicken ovalbumin, bovine fetuin, and horseradish peroxidase (HRP) TOF/TOF Printed onto commercially available Nhydroxysuccinimide (NHS) -activated glass slides after deprotection. Glycans interrogated using plant lectins and antibodies in sera from mice infected with Schistosoma mansoni (Song et al., 2009e) N-glycan clusters TOF (DHB) Biantennary and high-mannose N-glycans linked to non-reducing terminus of Man 3 GlcNAc 2 core, plus biotin (Song et al., 2009f) Thiol-terminated nonamannoside TOF Coupling of a thiol-terminated mannoside to maleimide-functionalized glass surfaces derived from γ -aminopropyl silane slides (Dietrich et al., 2010) Various oligosaccharides derivatized with 4-(2aminoethyl)aniline by reductive amination Reagent has amine groups at both ends allowing the modified carbohydrates to be covalently attached to an amino-reactive NHS-activated glass surface by formation of stable amide bonds (Seo et al., 2010) discussed in the paper. Finally, the method was validated by the determination of the chondroitin sulfate content in samples of known concentration as well as in enzymatically digested bovine nasal cartilage and compared with two further established methods of chondroitin sulfate determination (the carbazole and alcian blue methods) . Positive ion fragmentation of carbohydrates is fairly well understood with two types of cleavage, glycosidic cleavage that occurs between the sugar rings and involve hydrogen migration and cross-ring cleavages that involve the rupture of two of the bonds forming the rings. Glycosidic cleavages revealing sequence information predominate in positive ion spectra whereas negative ion spectra frequently contain very abundant cross-ring product ions that provide information on linkage. The nomenclature introduced by Domon and Costello (1988) is universally used to describe the ions. The stabilities of glycosyl bond linkages in various carbohydrates have been investigated by computational calculations to find general rules of fragmentation of sodiated oligosaccharides . The calculations revealed that a-glucose, a-galactose, b-mannose, a-fucose, b-GlcNAc and b-GalNAc linkages were cleaved more easily than b-glucose, b-galactose, and a-mannose linkages because the transition states of the former were stabilized by the anomeric effect. The 1-6 linkage was more stable than the others whereas the sialyl bond was the most labile of all the linkages investigated. Comparison of activation energies and binding affinities to the sodium cation revealed an increase in activation energy in proportion to the increment in binding affinity. The calculated stabilities of glycosyl bonds were: a-Man (Mana1 ! 3, 4 or 6Man ) > a-NeuNAc (NeuNAca2 ! 3 or 6); this result was close to the experimentally deduced trend. In-source decay frequently accompanies ionization of permethylated glycans. Although the presence of fragments can lead to complex spectra, they can also be used to obtain pseudo-MS 3 spectra. Smargiasso and De Pauw (2010) have investigated matrices and conditions for optimal production of such ions and have concluded that DHB was the most versatile matrix; the presence or absence of ISD fragments could be controlled, depending on the location of the laser shots. Spectra obtained from the center of DHB targets produced mainly [MþNa] þ ions that did not yield ISD fragments, whereas spectra from the crystals surrounding the target yielded mainly [MþH] þ ions that fragmented readily. 9-AA (6/18), on the other hand, formed homogeneous matrix spots and did not induce ISD. Soltwisch and Dreisewerd (2010) have noted that collisional cooling in an orthogonal TOF mass spectrometer stabilizes fragment ions that are generated in-source and that by varying the buffer gas pressure, production of ISD and post-source (PSD)-type ions could be varied. Under high-pressure conditions, ISD-type fragments of O-linked glycosylated peptides were generated that retain the glycan. PSD fragments, on the other hand, showed partial loss of the glycan from y-type peptide fragments. The detailed positive ion PSD and ISD fragmentation of deprotonated D-ribose (1/1) and D-fructose (1/16) has been studied with the aid of the isotopically labeled analogues, [1-13 C]-D-ribose, [5-13 C]-D-ribose and [C-1-2 H]-ribose (Bald et al., 2009) . The fragmentation was compared with fragmentation through dissociative electron attachment (DEA). Fragmentations of deprotonated monosaccharides formed in the MALDI process, as well as their transient molecular anions formed upon electron attachment, were characterized by loss of different numbers of H 2 O and CH 2 O units. Two different fragmentation pathways leading to cross-ring cleavage were identified. Metastable decay of deprotonated D-ribose proceeded either via an X-type cleavage yielding fragment anions at m/z ¼ 119, 100, and 89 (dominant ion, C 3 H 5 O 3 ), or via an A-type cleavage resulting in m/z ¼ 89, 77 and 71. This result is in contrast to previous CID studies where only A-type cross-ring cleavage was proposed. It was found that the heavier fragment anions at m/z ¼ 119 and m/z ¼ 100 generated via metastable decay exclusively arise from an X-type cleavage whereas the smaller fragment anions at m/z ¼ 89 and 71 arise predominantly from an A-type cleavage. A fast and early metastable cross-ring cleavage of deprotonated D-ribose observed in ISD was dominated by X-type cleavage leading mainly to m/z ¼ 100 and 71; the latter ion is not the same as that found in PSD. For DEA of D-ribose, a sequential dissociation was identified that included metastable decay of the dehydrogenated molecular anion leading to m/z ¼ 89. The most dominant fragment ions in DEA were due to faster decompositions occurring within several hundred nanoseconds (as in ISD) and, thus, sequential reactions including an initial dehydrogenation could be excluded. Several oligosaccharides (aldoses) and oligosaccharide alditols derived from agaroses, kand i-carrageenans obtained by hydrolysis of agaroses and carrageenans, two important types of red seaweed polysaccharides, have been used as model compounds to study prompt (ISD) and PSD fragmentation. Sulfated alditols gave [MÀH] À ions in negative-ion mode together with prompt fragment ions produced mainly by desulfation. Sulfated aldoses gave mainly prompt fragmentation ions (C-cleavages and desulfation). Non-sulfated aldoses and alditols only gave ions ([MþNa] þ ) in positive ion mode with no prompt fragmentation. The aldoses yielded cross-ring fragmentation in the PSD mode. Several different matrices (DHB, norharmane (1/35), ferulic acid (1/41) and the ionic liquid matrices DHB/acid-n-butylamine and ferulic acid/n-butylamine) were investigated; the best results were obtained with DHB and nor-harmane (Fatema et al., 2010) . CID on TOF/TOF and magnetic sector instruments have been compared with several types of biomolecule . The sector instruments produce high-energy collisions (8-20 keV) yielding more structural information from many compounds than instruments producing only low energy (1 keV) collisions. The case with different TOF/TOF instruments is less clear-cut because the collision energy is spread over a wide range. Fewer differences were seen with carbohydrates than with some other compound types because most fragmentations (except formation of X-type cross-ring cleavages) occur at low energies. High-energy fragmentation in positive ion mode generally enhanced the relative abundance of cross-ring cleavage fragments, particularly X-type ions and has been used with HPLC (offline) to provide a powerful method for glycoprotein analysis . Full experimental details are given in the paper. Experimental details for obtaining infrared multiphoton dissociation (IRMPD) spectra from carbohydrates have been described . The technique offers the advantage that, because both parent and product ions can absorb IR photons, the spectra can be richer in information than those obtained by CID. In the same publication, the authors discuss sustained off-resonance irradiation-collision-induced dissociation (SORI-CID) implemented with a MALDI-FT-ICR mass spectrometer which produced similar spectra to IRMPD. Experimental details are described. SORI-CID with MALDI-FT-ICR has also been applied to MUC-type O-glycans . The 157 nm photodissociation of N-linked glycopeptides has been investigated in a modified MALDI TOF/FOF instrument by irradiating the ions within the collision cell. Singly charged glycopeptide ions from horseradish peroxidase (HRP) yielded abundant peptide and glycan fragments. The peptide fragments included a series of x-, y-, v-, and w-ions with the glycan remaining intact. These ions provide information about the peptide sequence and the glycosylation site. The glycan fragmented to give both glycosidic fragments and abundant cross-ring fragments that were not observed in low-energy CID spectra. Doubly charged glycopeptides generated by nanospray in a linear IT mass spectrometer also yielded peptide and glycan fragments. However, the former were dominated by low-energy fragments such as b-and y-type ions while the glycan was primarily cleaved at glycosidic bonds (Zhang & Reilly, 2009 ). MALDI-LIFT-TOF-MS/MS and ESI tandem mass spectrometry (ESI-IT-MS/MS n ) have been used for the characterization of free oligosialic acids and those derivatized with DMB, as well as partially O-acetylated derivatives. Electrospray required the acids to be lactonized but the larger lactones could only be detected by MALDI-TOF. The fragmentation spectra were dominated by simultaneous cleavage of glycosidic linkages and the corresponding lactone ring, whereas classical cross-ring fragments were of minor abundance. However, the combined use of the two different types of fragmentation analysis allowed a sensitive and detailed characterization of both short-and longchained species. Furthermore, oxidation of the nonreducing end sugar moiety enabled sequence determination and localization of acetylated and nonacetylated sialic acid residues (Galuska et al., 2010a) . The effect of the pressure of cooling gas in the ion source of an orthogonal-TOF MS has a strong influence on the extent of analyte ion fragmentation. Soltwisch et al. (2009) have investi-gated the effect of this parameter on peptide and oligosaccharide fragmentation using substance P and the milk sugar, LNFP-II (52), respectively in both UV-and/or IR-MALDI. A range of pressures, from 0.05 to 1.8 mbar was used in combination with seven different buffer gases (He, Ne, Ar, N 2 , CO 2 , CH 4 , and isobutane). The influence of the ion extraction voltage on the analyte fragmentation was also investigated for a selected set of gas parameters. Fragment ions exhibited characteristic fragment yield-pressure dependencies that were classified into three groups. For LNFP-II, the yield of molecular ions rose with pressure until at the higher pressures, it was similar to that from substance P. The authors speculated that the consistently lower ion yields reported from oligosaccharides could be attributed to the generally low pressures used when recording their spectra. With respect to fragmentation, glycosidic fragment ions (termed Type I) ions dominated the spectra at low pressure but their relative abundance fell dramatically as the pressure rose. The ions resembled species that are also found in MALDI-PSD MS. The abundance of type II ions, which resembled typical ISD fragments, and consisted mainly of cross-ring products, rose with pressure, probably because of a reduction in the secondary fragmentation process that resulted in loss of fucose. A few other ions, termed type III ions did not show such dramatic changes with pressure. Comparing the yields of fragmentation for the different buffer gases revealed a correlation between their internal degrees of freedom and their collisional cooling efficiency. Changing the buffer gas pressure and/or extraction field provided an easy means to influence analyte ion fragmentation and to switch from the primary production of one type of fragment species to another. Hexose rearrangements of protonated molecules of N-glycopeptides and reductively aminated N-glycans have been observed by MALDI-TOF/TOF-MS/MS and ESI-IT-MS/MS (Wuhrer, Koeleman, & Deelder, 2009b) . Fragmentation of proton adducts of 2-AB and 2-AA-labeled high-mannose N-glycans from RNaseB resulted in transfer of one to five Hex residues to the fluorescently labeled innermost GlcNAc residue. Glycopeptides from various biological sources containing high-mannose glycans were likewise shown to undergo Hex rearrangement reactions, resulting in migration of one or two Hex moieties to the chitobiose core. Tryptic Fc-glycopeptides from IgG peptides containing biantennary N-glycans were also shown to undergo Hex rearrangements. Such rearrangement products can cause major problems with the interpretation of unknown glycans but, fortunately, do not appear to occur from [MþNa] þ ions, the major ions in MALDI spectra of most neutral carbohydrates. The use of computer software for analyzing carbohydrate spectra is not as advanced as that for proteins; nevertheless, many investigators have developed software for spectral interpretation, composition calculations, and have constructed databases, usually for specific glycan types. Much of this software is applicable to spectra generated from the majority of common ion sources. A good source of information is the book Bioinformatics for Glycobiology and Glycomics: An introduction by the late Claus-Wilhelm von der Lieth, Luetteke, and Frank (2009) . In addition, various tools for glycomics and available databases are covered in a comprehensive review by Frank and Schloissnig (2010) . The simplest of these algorithms calculates compositions from m/z values. Although such software is extremely valuable, a composition is not a structure and such programs should not be used as the basis of labeling spectra unless the proposed structures are confirmed by suitable techniques. One such tool reported in the review period is called lipID and calculates molecular compositions for glycerophospholipids, GSLs, fatty acids and small oligosaccharides from m/z values entered as single values or as mass lists. The user-extendable software is a Microsoft Excel Add-In developed using Visual Basic for Applications and is compatible with all Versions of MS Excel since MS Excel 97 (Hübner, Crone, & Lindner, 2009) . Kronewitter et al. (2009) have constructed a library of possible N-glycan masses by successive dismantling of tetraantennary hybrid and high-mannose glycans. These calculations gave the possible masses that would be expected in a glycan mixture. Three hundred thirty one distinct neutral compositions were obtained but many of these will represent several isomeric glycans. The smallest mass difference that was observed was 0.37 Da. However, many of the masses coincided with isotope peaks from ions of different compositions meaning that, without deisotoping, a resolution of at least 12,500 would be needed to resolve all peaks. The theoretical masses were matched against measured masses from N-glycans released from human serum glycoproteins and 78 discrete compositions were detected. In a similar way, an in silico glycan database of possible Nglycan compositions has been constructed by addition of known monosaccharide residues, such as those in a Neu5Ac-Gal-GlcNAc antenna, to the common trimannosyl chitobiose core. The derived masses were then matched to the experimental mass and the software, named Glyquest, predicted compositions and possible structures. Next, it calculated possible glycosidic fragments from the proposed structures, matched these to the experimental mass spectrum and constructed a spectrum labeled with the proposed structures (Gao, 2009) . The software could also be applied to glycans containing fluorescent labels such as 2-AB but was not applicable to glycopeptides with unknown modifications. However, as with much of the software developed for this work, the detailed structural details such as the linkage of each monosaccharide are not available and the software must be regarded as a guide to the total structure. A similar "branchand-bound" algorithm developed by Peltoniemi, Joenväärä, and Renkonen (2009) starts with the trimannosyl-chitobiose core and then constructs N-type glycans in an iterative process until the target carbohydrate composition is reached. The algorithm identified several glycans from TF and human serum samples. GlycoSpectrumScan is a web-based tool that identifies the glycoheterogeneity on a peptide from mass spectrometric data. Two experimental data sets are required as inputs: (1) oligosaccharide compositions of the N-and/or O-linked glycans present in the sample and (2) in silico derived peptide masses of proteolytically digested proteins with a potential number of Nand/or O-glycosylation sites. GlycoSpectrumScan uses MS rather than MS/MS data, to identify glycopeptides and determine the relative distribution of N-and O-glycoforms at each site. It can be used to assign monosaccharide compositions on glycopeptides with either single or multiple glycosylation sites. The algorithm allows the input of raw mass data, including multiply charged ions, making it applicable for both ESI and MALDI data from all mass spectrometer platforms. Low resolution data from, for example, ITs are heavily smoothed to yield the average mass whereas data from high resolution instruments receive a milder smooth and deisotoping to give the monoisotopic mass. The software was used to characterize the N-and O-linked glycopeptides from human secretory IgA (sIgA), consisting of secretory component (7 N-linked sites), IgA1 (2 N-linked, 5 O-linked sites), IgA2 (4 N-linked sites) and the J-chain (1 N-linked site). GlycoSpectrumScan is freely available at www.glycospectrumscan.org (Deshpande et al., 2010) . Prediction of glycosylation sites is another area of software development. A program that predicts N-and O-glycosylation sites based on local information, general protein information, sub-cellular localization and binding specificity of glycosyltransferases has been developed and was claimed to be about 90% accurate (Sasaki, Nagamine, & Sakakibara, 2009a) . However, as with all predictive programs that are not 100% accurate, results should only be taken as a guide for designing appropriate location experiments. Software that attempts to predict structures from spectra is possibly the most active area in computer applications. SimGlycan 1 is one such tool (Apte & Meitei, 2010) . The software accepts raw or standard experimental MS data files, matches them with its own database of theoretical fragments and generates a list of probable candidate structures. Each structure is scored to reflect how closely it matches the experimental data. The software also predicts novel glycan structures by drawing a glycan and mapping it onto the experimental spectrum. Other biological information is also available for easy reference. The program can be downloaded from http://www.premierbiosoft. com/glycan/index.html. Another software platform for carbohydrate assignment is SysBioWare, developed by Vakhrushev, Dadimov, and Peter-Katalinić (2009) and designed to work directly from raw MS data. The data are first imported into the spectrum browser, baseline corrected and denoized. Peak detection is based on shape matching and the software detects monoisotopic m/z values and charge states. A biological filter is used during compositional analysis of the monoisotopic ions. The software was successfully tested with human urine. SysBioWare, a software platform developed for MS data evaluation in glycomics, has been applied to the interpretation of spectra from human serum GSLs. The masses of predicted ions arising from cleavages in the glycan and the ceramide moieties were calculated, thus enabling structural characterization of both entities. The calculated masses were then used to match with those in the spectra for structural identification . Böcker, Kehr, and Rasche Böcker (2009) have presented an algorithm for calculating glycan structures from tandem mass spectra. Twenty-four spectra (of [MþH] þ ions) of 2-AP-labeled N-glycans obtained from batroxobin (from Bothrops moojeni venom) were used as test compounds, The spectra were measured with a TOF/TOF instrument with a MALDI ion source and the algorithm rapidly predicted possible topologies. Biological restraints needed to be used to limit the predictions to reasonable structures. Goldberg et al. (2009) have compared three algorithms, "Max Subgraph," "Parsimony," and "RandomWalk" that make inferences about glycan synthesis from biological knowledge for their ability to assign structures from 71 single-MS spectra from a variety of tissues and organisms, containing more than 2,800 manually annotated peaks. Max Subgraph behaved better than the other two but only uniquely assigned the correct structure to about half of the peaks in 41 out of the 71 spectra. A computer model that predicts N-linked glycan profiles based on cellular enzyme activities has been developed (Krambeck et al., 2009) . The paper describes the expansion of a previously developed detailed model for N-linked glycosylation (Krambeck & Betenbaugh, 2005) with the further application to analyze MALDI-TOF mass spectra of human N-glycans. The glycosylation reaction network is automatically generated by the model, based on the reaction specificities of the glycosylation enzymes and allows prediction of the complete glycan profile and its abundances for any set of assumed enzyme concentrations and reaction rate parameters. A predicted mass spectrum of model-calculated glycan profiles is obtained and enzyme concentrations are adjusted to bring the theoretically calculated mass spectrum into agreement with that obtained experimentally. The result is a complete characterization of a measured glycan mass spectrum containing hundreds of masses in terms of the activities of 19 enzymes. In addition, a complete annotation of the mass spectrum in terms of glycan structure is produced, including the proportions of isomers within each peak. The method was applied to mass spectrometric data obtained from normal human monocytes and monocytic leukemia (THP1) cells. A kinetic model originally developed for the prediction of peptide CID spectra has been extended to predict the CID spectra of N-glycopeptides. The model was trained with 1831 CID spectra obtained with an ion trap and was able to predict CID spectra with excellent accuracy in ion intensities for N-glycopeptides up to 8,000 Da in mass. The program is said to be capable of predicting up to 524 common glycoforms including high-mannose, hybrid and complex N-glycans with two to four antennae (Zhang & Shah, 2010) . Spencer et al. (2010) have devised a computational approach to predict the fine structure and patterns of domain organization of heparan sulfate (HS). Analysis uses chemical composition data obtained after complete and partial enzymatic digestion of mixtures of HS chains and produces populations of theoretical HS chains with structures that meet both biosynthesis and enzyme degradation rules. The program was used to analyze HS from various cell types and good agreement was found between experimental data and computer predictions. GlycoViewer (http://www.systemsbiology.org.au/glycoviewer) is a web-based tool that can visualize, summarize, and compare sets of glycan structures. It takes as its input a list of glycan structures in International Union of Pure and Applied Chemistry (IUPAC) format or glycan structures constructed with a sugar structure builder. The output is a graphic, which summarizes all salient features of the glycans according to features such as the nature and length of any chains and the types of terminal epitopes. The tool can summarize several hundred glycan structures in a single figure. The report contains an example of use of the tool for analysis of normal and disease associated glycans from the human glycoproteome (Joshi et al., 2010) . Several glycan databases are available. The kyoto encyclopedia of genes and genomes (KEGG) GLYCAN databases contain useful information on glycan structures and metabolic pathways (Hashimoto & Kanehisa, 2009) and data mining the Protein Data Bank for glycol-related data using the GLYCOSCIENCES. de internet portal has been discussed in Methods in Molecular Biology (Lütteke & von der Lieth, 2009) . Ito et al. (2010a) have synthesized N-and O-linked glycan libraries (named Glycan Mass Spectral Database, GMDB) and constructed a library of their positive ion MS 2 , MS 3 and MS 4 fragmentation spectra. N-Glycans were in the form of their 2-AP derivatives whereas Oglycans that were released by b-elimination were not. The library was said to be accessible on-line at http://riodbdev.ibase. aist.go.jp/rcmg/glycodb/Ms (However, attempts by the reviewer to connect to the site have failed.) It can be searched either by MW of glycan composition in terms of isobaric monosaccharides and instructions on how to use the software are given in the paper. Although such databases and tools for glycomics are readily available on the web, these have, until now, been isolated. This unfavorable situation has been discussed (von der Lieth, 2007) and has been largely overcome by Ranzinger et al. (2009) who have developed GlycomeDB, a meta-database for public carbohydrate sequences. At the time of publication (2009) it contained 35,056 unique structures in GlycoCT (www. glycome-db.org) encoding, referencing more than 100,000 external records from 1,845 different taxonomic sources. A user-friendly, web-based graphical interface has been developed which allows taxonomic and structural data to be entered and searched. The structural search possibilities include substructure search, similarity search, and maximum common substructure. A novel search refinement mechanism allows the assembly of complex queries. With GlycomeDB, it is now possible to use a single portal to access all digitally encoded, public structural data in glycomics and to perform complex queries with the help of a web-based user interface. A list of databases is given in Aoki-Kinoshita (2010). N-and O-glycan structures are usually depicted with small cartoons in which each constituent monosaccharide is shown by a symbol. Unfortunately, there is no consensus on which symbol to use for any particular monosaccharide with most investigators preferring to develop their own system. Several years ago, the Consortium for Functional Glycomics (CFG) attempted to redress the problem with the introduction of a system that has since been adopted by several laboratories. Unfortunately, this system has several major drawbacks: (a) it does not diagrammatically show linkage or anomericity and (b) it uses color to differentiate hexoses, thus causing problems when structures are printed in black and white and making the system difficult to use with pen or pencil on paper. A new system that overcomes these problems has recently been introduced (Harvey et al., 2009c) . Monosaccharides are shown as shapes with various additions to indicate functional groups (e.g. an inclusive dot to indicate a deoxy-sugar and a filled shape to code for an N-acetyl sugar). Linkage is shown by the angle of the lines linking the sugar symbols and anomericity is shown by the type of line (full for a b-bond and broken for an a-link). Examples can be seen below. Although color is not used to define monosaccharides, the CFG colors can be used with the Oxford symbols. Unfortunately, color was omitted from the table of symbols in the original article but was published later as an erratum (Harvey et al., 2009b) . The article received comments from the authors of the CFG system (Varki et al., 2009) and discussion is continuing. This scheme and others are compared in a review by Frank and Schloissnig (2010) . Two tools for displaying N-glycans found in the mammalian CHO cell line have been developed (McDonald et al., 2010) . Both take as input the 9-digit identifier devised by Krambeck and Betenbaugh (2005) that uniquely defines each structure assuming the existence of the trimannosyl-chitobiose core. The first of these tools, GlycoForm, is designed to display a single structure from an identifier entered by the user. The display is updated in real time, using symbols for the sugar residues, or in text-only form. The two symbol sets discussed above are used, the symbols and layout devised by the CFG http://www. functionalglycomics.org/static/consortium/Nomenclature.shtml or the alternative "Oxford" system used by Glycobase, a relation database of 2-AB labeled N-glycans (Campbell et al., 2008) . However, although GlycoForm can display structures using the Oxford system, unfortunately, it does not display the correct linkage information that is inherent to the full Oxford system. In addition, GlycoForm can display the name of the glycan as used by Glycobase. GlycoBase formalism does not yet handle Nacetyllactosamine (NAcLac, 53) repeating units and is, therefore, currently limited to structures with one Gal residue per branch. Structures can be added to a library, which is recorded in a preference file and loaded automatically. Individual structures can be saved as image files either Portable Network Graphics (PNG), JPEG or Windows Bitmap (BMP) formats. The second program, Glycologue, reads a file containing columnar data of nine-digit codes, which can be displayed on-screen and printed at high resolution. Both programs, for Windows, Mac OS X and Linux x86 GTK can be downloaded from http://www.boxer.tcd. ie/gf. A major problem with the analysis of monosaccharides or small oligosaccharides by MALDI is the presence of very abundant ions from matrices such as DHB in the low mass region of the spectra. Various methods for overcoming this disadvantage are discussed above. Nevertheless, MALDI with conventional matrices has produced results and the technique works well for the larger oligosaccharides. Thus, oligosaccharides from dextran, alginate, hyaluronan and chondroitin sulfate have been characterized by MALDI-TOF MS directly from a TLC plate after soaking it in the DHB matrix. The plate had a metal backing to ensure electrical contact. The TLC solvent system was n-butanol/formic acid/water (3:4:1, v/v/v). It was found that the high content of formic acid caused few problems but was responsible for partial formylation of glycosaminoglycans (GAGs) and minor N-acetyl loss from hyaluronan and chondroitin sulfate . A comparative study of MALDI and a new technique, electrospray droplet impact secondary ion mass spectrometry (EDI/SIMS) has been applied directly to fruits such as bananas, apples, grapes and strawberries. The major constituents, fructose (1/16), glucose (1/4), sucrose (6) and organic acids gave abundant [M þ K] þ ions positive mode and CF 3 COO À adducts in negative mode (the CF 3 COO À ions came from CF 3 COOH in the ESI spray. These negative ion spectra were almost free of background ions. MALDI from DHB, on the other hand, although producing positive ions gave virtually no ionization in negative ion mode (Asakawa and Hiraoka, 2010) . Reviews of the analysis of polysaccharides are listed in Table 5 . Large polysaccharides need to be hydrolyzed, often enzymatically but sometimes chemically, to smaller fragments before they are amenable to MALDI analysis. One chemical method involves the selective cleavage of the Rhap-(1 ! 4)-a-GalAp linkage in rhamnogalacturonans. Enzymic cleavage of this linkage is often ineffective, especially in highly branched rhamnogalacturonans but Deng et al. (2009) have developed an improved chemical fragmentation method based on belimination of esterified 4-linked galacturonic acid (GalpA, 54) residues that overcomes the problem. At least 85% of the carboxyl groups of the GalpA residues in A. thaliana seed mucilage were converted to methyl or hydroxypropyl esters and b-elimination was found to be more extensive with hydroxypropyl-esterified than with methyl-esterified rhamnogalacturonans. The non-reducing 4-deoxy-b-l-threo-hex-4-enepyranosyluronic acid (55) residue formed by the b-elimination reaction was removed by treatment with aqueous N-bromosuccinimide. This method was used to fragment the branched rhamnogalacturonan from peppergrass seed mucilage with product characterization by MALDI-TOF MS, glycosyl residue composition analysis, and 1 and 2D NMR spectroscopy. The results showed that the most abundant low-MW fragments contained a backbone rhamnose (1/10) residue substituted at O-4 with a single side-chain, and suggest that peppergrass seed mucilage rhamnogalacturonans is composed mainly of the Another chemical method for analysis of rhamnogalacturonan II makes use of mild acid hydrolysis to hydrolyze the acidlabile monosaccharides 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo, 1/13), 3-Deoxy-D-lyxo-2-heptulosonic acid (Dha) and apiose (Api, 56) at the branchpoint between the sidechains and the oligogalacturonide backbone to release short polysaccharide chains that were analyzed by ESI and MALDI-TOF MS (Séveno et al., 2009) . The method was optimized using citrus pectin and then applied to other plant species. Experimental details for examination of extracellular polysaccharides (EPSs) from plants following digestion with a variety of endoglycosiodases and MALDI-TOF analysis from DHB has been reported by Günl, Gille, and Pauly (2010) . Other examples are listed in Tables 6 (plants) and 7 (bacteria). Ionization efficiencies of cyclodextrins and corresponding linear compounds (maltohexaose and maltoheptaose) have been compared together with differences in the ionization efficiencies of aand b-cyclodextrins (4/6) . Alkali metal salts of Li þ , Na þ , K þ , and Cs þ were used as the cationizing agents to enhance the ionization efficiency. Relative ion intensities of the cyclodextrins were much larger than those of the linear carbohydrates and the difference showed an increasing trend with the size of the alkali metal cation. b-Cyclodextrin had higher ionization efficiency than a-cyclodextrin (4/24) and the difference increased with increasing size of the alkali metal cation. The ionization efficiency was also found to be affected by the counter anions. The higher ionization efficiencies of cyclodextrins were explained with the number of coordination sites and the binding energies. Analysis of milk oligosaccharides appears to be receiving increasing attention. Reviews of mass spectrometric methods for their analysis have been published by Niñonuevo and Lebrilla (2009) , Kolarich and Packer (2010) and Urashima et al. (2009) . Wu et al. (2010b) have developed an annotated library of neutral human milk oligosaccharides with characterization by HPLC, MALDI-FT-ICR MS and exoglycosidase digestion. Pyrene labeling (Amano et al., 2009a) has been used by Amano et al. (2009b) to enable neutral carbohydrates from human milk to be observed by negative ion MALDI-TOF MS n . The neutral oligosaccharides from the milk of a woman (blood type A, Le bþ ) were obtained by anion-exchange column chromatography after the removal of lipids and proteins. Further fractionation was performed by means of Aleuria aurantia lectin-Sepharose column chromatography and reversed-phase HPLC after labeling. Twenty-two oligosaccharides with decaose cores were identified and, of these 21 had novel structures. LoCascio et al. (2009) have published a method for measuring the consumption of human milk oligosaccharides by 12 strains of Bifidobacteria. Oligosaccharides were quantified with deuterated and reduced oligosaccharide standards that were added after bacterial growth and results were processed with inhouse software called Glycolyzer after removal of contributions from 13 C isotopes. High growth was found for Bifidobacterium longum biovar infantis strains, which consumed nearly all available substrates, while other bifidobacterial strains showed low or only moderate growth ability. Other examples of the use of MALDI analysis of milk glycans are listed in Table 8 . Glycoproteins and their attached N-and O-glycans is possibly the largest group of compounds that have been analyzed by MALDI, catalyzed largely by developments in the biotechnology industry. Analysis of these compounds has been reviewed by many authors (Table 9) . N-glycans are normally attached to an asparagine residue in a Asn-(Xxx)Ser-(Thr) consensus sequence where Xxx is any amino acid (Xxx) except proline However, Asn-linked N-glycans have recently been found at the 0.5-2.0% level on a non-consensus amino acid sequence (TVSWN 162 SGAL) in the C H 1 domain of human antibodies and on IgG1 (Valliere-Douglass et al., 2009b). Many investigators have published methods for glycoprotein enrichment. Solid-phase glycan/glycoprotein capturing methods have become popular in recent years and some of these have been highlighted in an article by . (Kooy et al., 2009) Upis ceramboides (Alaskan beetle) Xylomannan First identification of a nonprotein anti-freeze compound (Walters et al., 2010) Commercial Chitosan oligosaccharides Analysis of commercial samples and preparation of sample with GlcNAc 5-12 Not stated but has been found in Cryptococcus neoformans Human macrophage activation shown to be triggered by chitotriosidase-mediated chitin and chitosan degradation (Gorzelanny et al., 2010) 1 Format (not all items present): MALDI method (matrix), compounds run (derivative), other methods. Enrichment strategies for glycopeptides based on lectinaffinity chromatography and polysaccharide hydrophilic affinity physicochemical chromatography have been discussed by Ito, Hayama, and Hirabayashi (2009b) . The combined use of these techniques effectively removes non-glycosylated peptides. The ability of boronic acids to form cyclic derivatives with the cis-dihydroxy groups present in most glycans has been extensively used. Thus, 3-aminophenylboronic (APB) acid (5/42)functionalized beads, mesoporous silica, and nanodiamonds have been developed to enrich glycosylated peptides and proteins but the direct immobilization of the APB group was found to be insufficient to suppress nonspecific adsorption/ adhesion. Consequently Jang et al. (2009b) have designed a selfassembled monolayer (SAM)-based plate, which contained a spacer group such as oligo(ethylene glycol) to reduce the nonspecific adsorption/adhesion, for direct detection of glycoproteins after affinity-capture (or enrichment) on the plate. The utility of the plate was demonstrated with model glycoproteins such as ribonuclease G and TF. A two-stage glycopeptide enrichment technique using boronate-functionalized beads has been developed by Chen et al. (2010f) . Samples were incubated with the functionalized magnetic beads in slightly alkaline conditions at room temperature for about 1 hr with gentle shaking. The beads were washed and the enriched glycoproteins/peptides were eluted under acid conditions and dried in a Speed-Vac evaporator. The glycoproteins were then either dissolved in 50 mM ammonium bicarbonate and digested by Lys-C overnight or separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and the resulting gel-bands were digested in-gel by Lys-C overnight. The compounds were then ready for a second enrichment. Alternatively, digestions could be carried out with trypsin. Analysis was by MALDI-TOF. Boronic acid functionalized nanoparticles have also been used to concentrate antibodies by capturing the carbohydrates attached to the Fc region of IgG . Chalagalla and Sun (2010) have prepared a boronic acid-containing polymer capped with biotin (57) for linkage to a magnetic bead and used the product for glycan capture and Lin et al. (2010f) have constructed magnetic nanoparticles with immobilized APB acid for glycoprotein capture. "SnO 2 @Poly(HEMA-co-St-co-VPBA)" Core-shell nanoparticles containing boronic acid groups have been prepared by of copolymerization between 2hydroxyethyl methacrylate grafted (58) on SnO 2 nanoparticles, styrene, and 4-vinylphenylboronic acid (VPBA, 59). They have been used to extract tryptic peptides from HRP, bovine asiloTF and human serum glycoproteins. Analysis was by MALDI-MS/ MS using an AXIMA QIT instrument (Sheng, Xia, & Yan, 2010 Survey EPO and of several analytical methods including MALDI, chromatography 175 (Reichel & Gmeiner, 2010) Application of proteomics in biomarker discovery Mainly proteins and glycoproteins. A novel boronic acid functionalized mesoporous silica, which holds the attractive features of high surface area and large porosity has also been used to concentrate glycopeptides. In comparison to direct (traditional) analysis, this method was stated to enabled two orders of magnitude improvement in the detection limit of glycopeptides irrespective of the nature of the attached glycans . The same group has also synthesized boronic acid functionalized core-satellite composite nanoparticles that possess both the superparamagnetic properties of magnetic nanoparticles and the surface chemistry of AuNPs. Glycoproteins or glycopeptides could be obtained in high yield by use of a magnet. The composite nanoparticles were used to enrich glycosylated proteins from human colorectal cancer tissues for identification of N-glycosylation sites. In all, 194 unique glycosylation sites mapped to 155 different glycoproteins were identified, of which 165 sites (85.1%) were new. Boronic acid functionalized gold-coated Si wafers have been used as MALDI plates to isolate and enrich glycopeptides . This method was claimed to be beneficial for several reasons. Thus, solution transfer and eluting steps required in conventional enrichment strategies were not needed, thereby reducing sample loss. Secondly, the lower limits of detection of glycopeptides were said to have been increased by two orders of magnitude. Thirdly, non-specific bindings were not detected even when non-glycopeptides were 100 times more concentrated than glycopeptides. Furthermore, glycopeptides could be detected in the presence of 200 mM ammonium bicarbonate or the physiological buffer, PBS. In a related method (Tang et al., 2009a; Yao et al., 2009) , AuNPs were first spotted and sintered onto a stainless steel plate, then modified with 4-mercaptophenylboronic acid (60) to provide a porous substrate with a large surface for capturing glycopeptides from peptide mixtures. The captured peptides were then analyzed by MALDI-TOF MS simply by deposition of a DHB matrix. The technique enabled sample enrichment, washing and detection steps to be fulfilled on a single MALDI target plate. Well-characterized glycoproteins, such as HRP and asialofetuin, were employed as standards to investigate the enrichment efficiency. Fe 3 O 4 @C@Au magnetic microspheres functionalized with 4-mercaptophenylboronic acid have been synthesized by the same group (Qi et al., 2010) and successfully used for enrichment of glycoproteins and glycopeptides. A polyfunctional device has been constructed from Macroporous silica foam (MOSF) containing boronic acid (BMOSF) or amino groups (NH 2 -MOSF) and used to immobilize enzymes such as trypsin and selectively enrich glycopeptides. Use of the device considerably speeded up hydrolysis times as demonstrated with glycopeptides from HRP (Qian et al., 2010) . Tang et al. (2010a) have immobilized the lectin Con A on APB acid-functionalized magnetic nanoparticles using methyl a-D-mannopyranoside as a linker. The selective capturing ability of the Con A-modified nanoparticles was tested using standard glycoproteins and cell lysate of human hepatocelluar carcinoma cell line 7,703. Regeneration of the protein-immobilized nanoparticles could easily be performed by utilizing the reversible binding between the boronic acid and the sugar. ConA has also been used in conjunction with hollow fiber flow field-flow fractionation (HF5) to preconcentrate high mannose type Nlinked glycoproteins from bacterial lysates as exemplified by glycoproteins from Streptococcus pyogenes . The specificity of Datura stramonium agglutinin (DSA) for triand tetra-antennary glycans has been utilized to enrich human liver glycoproteins containing these larger glycans which were then separated and identified by SDS-PAGE followed by MALDI-TOF analysis (Sun et al., 2009b) . The performance of chromatographic columns consisting of agarose-bead-bound 3-aminophenyl boronic acid, agarosebound wheat-germ agglutinin (WGA) or a mixture of both compounds (boronic acid lectin affinity chromatography, BLAC) has been evaluated for glycoprotein enrichment using the model proteins of RNaseB and trypsin inhibitor in the presence of the non-glycosylated proteins, myoglobin (neutral) and lysozyme (basic) over a wide temperature range (5-65˚C). The results showed that glycoaffinity micropartitioning at 25˚C provided the highest recovery rate for glycoprotein enrichment. A large amount of lysozyme was present in the elution fractions of the 3-aminophenyl boronic acid-containing micropartitioning columns due to an ion-exchange mechanism occurring between the positively charged protein and the negatively charged stationary phase. At 65˚C, nonspecific interactions with the agarose carrier prevailed, evidenced by the presence of myoglobin in the eluate (Olajos et al., 2010) . A novel boronate affinity monolith, poly-(3-acrylamidophenylboronic acid-co-ethylene dimethacrylate) (61) has been prepared in 530 mm capillaries by a one-step in situ polymerization procedure . The monolith was used to separate glycopeptides from peptides produced from HRP and to separate this glycosylated protein from non-glycosylated bovine serum albumin (BSA). The MALDI-TOF spectrum of the HRP peptides showed little evidence of the presence of glycopeptides before passage through the capillary but revealed abundant glycopeptide ions after treatment. Mass Spectrometry Reviews DOI 10.1002/mas b. Other solid-phase methods Titanium dioxide (TiO 2 ) microspheres, synthesized by a sol-gel method, have a high affinity for the acid groups of sialic acids and peptides. They have successfully been used for simultaneous enrichment of glycopeptides and phosphopeptides from, for example bovine RNaseB and human IgG . Detection was by ESI but the method would be equally applicable to MALDI-TOF analysis. Mysling et al. (2010) have used ZIC-HILIC in a microcolumn format for SPE and glycoprotein enrichment involving trifluoroacetic acid (TFA) ion pairing to increase the hydrophilicity difference between glycopeptides and non-glycosylated peptides. Three mobile phases were investigated: 2% formic acid, 0.1% TFA and 1% TFA all containing 80% acetonitrile and experiments were conducted on single glycoproteins, a five-glycoprotein mixture and depleted plasma. The presence of TFA, particularly at the 1% level, in the mobile phase significantly improved the glycopeptide enrichment (3.7-fold) as evaluated by MALDI-TOF MS and RP-LC-ESI-MS/MS. Four types of hydrazine functionalized carboxyl and epoxysilanized magnetic particles (HFMP) have been developed by Sun et al. (2010a) for isolation of glycopeptides. Particles prepared by adipic dihydrazide functionalization from carboxylsilanized magnetic particles yielded the maximum capture capacity. The method was verified by successful isolation of all formerly glycosylated peptides from standard glycoproteins (fetuin, RNaseB, and human serum albumin (HSA)) and by identification of their glycosylation sites. c. Other techniques MALDI-TOF MS has been used by Carvalho et al. (2009) to characterize the aand b-subunit of recombinant and pituitary glycoprotein hormones that have been separated by a new method of incubating the glycoproteins overnight with acetic acid (0.5-3.0 M) at 37˚C. a. Use of mass spectrometry to detect glycosylation of proteins A simple method to detect glycosylation is to measure the mass of a glycoprotein and then to repeat the measurement after incubation with PNGase F to remove the N-glycans. The method has been used to detect the presence of N-glycans in recombinant bovine CD38 expressed in Pichia pastoris (Muller-Steffner et al., 2010) . The molar masses of non-glycosylated (29,343 Da) and penta-glycosylated Thermomyces lanuginosus lipase (40,906) as measured by MALDI-TOF MS have confirmed glycosylation and shown that each glycan-moiety adds approximately 2,000 Da to the molar masses (Pinholt et al., 2010) . In another example, Cu/Zn superoxide dismutase monomer was determined to have a mass of 17,097 Da before deglycosylation and 15,871 Da afterwards giving a mass for the glycan of about 1,200 (Nedeva et al., 2009) . The difference between the sequence mass (33,768 Da) and measured mass of about 44,604 Da of the peroxidase from royal palm tree (Roystonea regia) together with the fact that the amino-acid sequence includes 12 possible N-glycosylation sites, suggests heavy glycosylation. Glycosylation sites were identified, in this case, by N-terminal sequencing and MALDI-TOF-MS analysis of tryptic peptides . b. Mass spectrometric detection of glycoforms of intact glycoproteins Although resolution of glycoforms by MALDI-TOF MS is generally inferior to that obtained by ESI, glycoproteins with masses in the region of 60 kDa, containing only a limited number of glycoforms have been resolved successfully as illustrated by the resolution of four glycoforms of antithrombin in a study of altered glycosylation causing antithrombin deficiency . Glycans appeared to be sialylated biantennary (residue mass 2,204 Da). Ogawa et al. (2009) have observed resolution of glycoforms of Stereum purpureum endopolygalacturonase I produced in P. pastoris (36.5 kDa). Three main ion peaks corresponding to the protein with the high-mannose glycans Man 8-10 GlcNAc 2 together with some minor unresolved ions were observed. c. Detection of glycosylation sites and site occupancy The most common method for detecting site occupancy is to utilize the conversion of the Asn to which the N-glycans are attached to aspartic acid (Asp) when the glycans are released with PNGase F. The increase by one mass unit is readily detected by MS. The method has been used, for example to confirm occupancy of six of the seven potential N-linked glycosylation sites in the envelope glycoprotein gp116 and three of the four potential sites in the gp64 protein of the yellow head virus from the Penaeus monodon shrimp (Soowannayan et al., 2010) . Periodate oxidation and hydrazide capture on a solid support have been used by Lewandrowski and Sickmann (2009) to study glycosylation sites in human platelet proteins. The bound glycoproteins were sufficiently stable to allow washing, following which the proteins were hydrolyzed. Glycopeptides remained bound to the solid support through the glycan moiety from where they were released with PNGase F and the glycosylation site was identified by means of the Asn to Asp conversion. A new method using tandem 18 O stable isotope labeling (TOSIL) to quantify the N-glycosylation site occupancy has been reported (Liu et al., 2010k) . Glycoproteins were digested with trypsin and PNGase F in the presence of either H 2 18 O or H 2 16 O. Three 18 O atoms were introduced into N-glycosylated peptides, two at the carboxyl terminus of all peptides and the third at the N-glycosylation site. The samples were mixed to give pairs of ions in the resulting MALDI or ESI spectra. A unique mass shift of 6 Da was produced by N-glycosylated peptide with a single glycosylation site, whereas non-glycosylated peptides produced an ion pair spaced by only 4 Da. Intensity ratios could be used to monitor site occupancy in various physiological and disease conditions. The method yielded good linearity within a 10-fold dynamic range with the correlation coefficient r 2 > 0.99. The standard deviation (SD) ranged from 0.06 to 0.21, for four glycopeptides from two model glycoproteins. The method was used to monitor glycoproteins in the sera from a patient with ovarian cancer and healthy individuals. Eighty-six N-glycosylation sites were quantified and N-glycosylation levels of 56 glycopeptides showed significant changes. A similar 18 O-labeling technique was used by Alvarez-Manilla (2010b) to identify N-glycosylation sites in Con-A-extracted glycopeptides from pluripotent murine embryonic stem cells. Glycopeptides rather than glycoproteins were extracted from tryptic digests to avoid false positives produced from non-glycosylated proteins that were bound to extracted glycoproteins if no digestion had been performed. Segu et al. (2010b) have published a method for detecting sites occupied by glycans carrying a fucose residue attached to the core. The method made use of the endoglycosidase M which, like PNGase F has a broad spectrum of activity with the exceptions that (a) it is not active on core-fucosylated glycans and (b) it shows reduced activity with larger glycans. The second exception was overcome by conducting incubations in the presence of sialidase, b-galactosidase and b-N-acetylhexosaminidase, which reduced the size of the glycans. Then, the results were compared with the products of digestions performed with PNGase F allowing the core-fucosylated sites to be determined. Analyses were by LC/MS but the technique would be equally applicable to MALDI-TOF analysis. d. Analysis of glycopeptides Because many glycoproteins are too large and heavily glycosylated for direct analysis by MS, much work is performed on derived glycopeptides, most commonly tryptic glycopeptides. Tryptic cleavage of glycoproteins is frequently hindered by steric hindrance imposed by the glycans but improvements can be made by the use of heat to increase the rate of proteolysis. Segu, Hammad, and Mechref (2010a) have used microwaveassisted enzymatic digestion to achieve higher sequence coverage of several model glycoproteins such as fetuin, TF, and fibrinogen. Efficient digestion was achieved in 15 min at an optimum temperature of 45˚C; there was no apparent loss or partial cleavage of the glycans. Signals from glycopeptides are often weak or absent from the spectra of mixed peptides and glycopeptides, a situation that can be improved by fractionation of the two compound classes. Wohlgemuth et al. (2009) have investigated several techniques and have shown that hydrophilic interaction chromatography (HILIC) chromatography with ZIC-HILIC and TSKgel Amide-80 are very specific at capturing glycopeptides from mixtures. Sialylated glycopeptides could also be enriched with TiO 2 . Capture using a hydrazide column resulted in lower recovery and involved a more complex enrichment scheme. A new material for glycopeptide concentration, termed "click maltose," has been synthesized by linking the alkynyl-derivatized maltose chain to the azide derivatized silica gel through click chemistry. Unlike the rigid structure of Sepharose, the saccharide chain of click maltose exhibits a certain amount of flexibility, which provides a sufficient number of hydroxyl groups for the effective formation of hydrogen bonds with the glycans attached to glycopeptides. The material was used to isolate glycopeptides from IgG, RNaseB, and AGP . Cellulose columns have also been used for concentration of glycopeptides (Snovida et al., 2010) . A method for detecting core-fucosylated (CF) glycoproteins for screening purposes has been reported by Jia et al. (2009) . After IgG depletion, fucosylated plasma proteins were enriched by use of Lens culinaris lectin and the bound glycoproteins were digested by trypsin. These compounds were enriched by use of a 3,000 Da cut-off filter, a procedure that also combines de-salting and concentration. The recovered glycopeptides were then treated with endoglycosidase F3, which specifically cleaves the glycosidic bond between the two proximal (core) GlcNAc residues and leaves the fucosyl-GlcNAc residues attached to the peptides. Four standard glycoproteins, apo-TF, fetuin, rhEPO, and RNaseB, was used to illustrate the method. In addition, a part of the untreated tryptic peptides was treated with PNGase F in order to locate the glycosylation site by the Asn to Asp conversion. Products were detected by MALDI-TOF. In a related method using an ion trap, a neutral loss scan for fucose (146 Da) was also used to detect fucosylation. The methods were applied to the detection of fucosylated glycoproteins in the plasma of healthy subjects and subjects with hepatocellular carcinoma. Over 100 fucosylated glycoproteins and attachment sites were identified, and over 10,000 mass spectra of CF glycopeptide were analyzed. A method termed the Sulfate Emerging method has been described for specifically extracting sulfated glycopeptides (Toyoda, Narimatsu, & Kameyama, 2009 ) from mixtures. The method overcomes the often negative contribution from other charged groups in the molecules. To accentuate the negative charge on the sulfate group, basic amino acids were eliminated and carboxylic acids were neutralized as follows: The protein was first hydrolyzed with trypsin and then the positively charged C-terminal lysines (Lyss) and arginines (Args) were eliminated by incubation with carboxypeptidase B. The negative charges of the carboxylic acid groups on the peptides were then neutralized by chemical modification with acetohydrazide using the recently reported quantitative modification of carboxyl groups in sialic acid using this reagent and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (43). The sulfated glycopeptides in the mixture were then captured by anion exchange resin with a basic buffer (pH 8.6) in which protonation of histidine residues was suppressed. Finally, the sulfated glycopeptides were eluted from the anion exchange resin by increasing the ionic strength of the elution buffer for analysis by MS. Rather than trypsin, pronase has been used as a nonselective enzyme to reduce the protein to a single amino acid (Asn) or short peptide attached to N-glycans. These compounds are generally smaller than those obtained from O-linked glycopeptides, probably because O-glycans lie closer to the peptide backbone than N-glycans and protect the polypeptide from enzymatic digestion. N-Glycans usually rise above the peptide backbone exposing the polypeptide to enzymatic digestion. Dodds et al. (2009) have described immobilized pronase which retains its activity after repeated use for at least 6 weeks. Use of negative ion detection has been reported by Nwosu et al. (2010) as providing distinct advantages over detection in positive ion mode for the detection of glycopeptides produced by pronase digestion. Analysis in positive ion mode, although most commonly used for glycopeptide characterization, is hampered by potential charge-induced fragmentation of the glycopeptides and poor detection of the glycopeptides carrying sialic acids. Furthermore, CID spectra of glycopeptides in the positive ion mode predominantly yields glycan fragments with minimal information on the peptide moiety. In the study by Nwosu et al., which employed bovine lactoferrin for detection of N-glycosylation and k-casein for O-glycosylation, 44 potential N-linked glycopeptides were detected in the positive ion mode whereas 61 potential N-linked glycopeptides were detected in negative ion mode. Analysis of k-casein, which contained mainly sialylated glycans, yielded improved results in negative mode. Experimental details for peptide mass fingerprinting and identification of glycosylation sites have been published by Wilson, Simpson, and Cooper-Liddell (2009) . Unlike the case with released glycans, where a relatively low mass accuracy measurement is usually sufficient to determine the composition in terms of the constituent monosaccharides, the situation is very different for measurements of glycopeptides. Desaire and Hua (2009) have examined the accuracy required and have concluded that in only a few cases can the mass accuracy provided by most commercial instruments be sufficient to unambiguously assign compositions to all glycopeptides in a mixture. e. N-Glycan release Once glycosylation sites have been determined, detailed structural analysis of the attached glycans is more conveniently carried out after releasing the glycans from the protein or peptide. Both chemical and enzymatic methods are available but although, in the past, chemical release with hydrazine was popular, most investigators now prefer enzymatic methods. i. Enzymatic release PNGase F. Peptide N-glycosidase F (PNGase F), an amidase, is the most popular enzyme for cleaving N-glycans from their Asn linkage site. It shows a broad range of substrate specificity with the exception that it does not release glycans bearing a fucose residue attached to position 3 of the reducingterminal GlcNAc; in these cases, PNGase A is appropriate. PNGase F cleaves the entire glycans, which are released as the corresponding glycosylamines. These compounds rapidly hydrolyze to the glycan with retention of the reducing terminus. This method is, thus, distinctly advantageous to techniques such as b-elimination, popular with O-glycans (see below) because this site can conveniently be used to attached tags for fluorescence or other detection methods. Wang et al. (2009i) have recently found that the activity of the enzyme towards denatured glycoproteins can be enhanced by removal of the Nterminal H1 helix from the enzyme. Bereman et al. (2009b) have studied methods for optimizing the release of glycans with this enzyme. Dialysis of plasma prior to incubation was found to have little or no effect. However, microwave-assisted glycan release was found to be beneficial; 20 min at 20˚C with approximately 250 W was found to give optimal results. Surprisingly, no protease digestion was required as needed with standard incubation methods, and it was found that an 18-hr incubation with no detergent (NP40) led to the greatest ion abundance of glycans from plasma glycoproteins. Data could be obtained in less than 1 day from raw plasma samples utilizing microwave irradiation. PNGase F-glycan release from human serum glycoproteins has been achieved in 10 min by using a constant microwave power of 20 W, giving a temperature of 44˚C (Kronewitter et al., 2010) . In this study, the glycans were recovered by SPE using a robotic liquid handler and examined by MALDI with an FT-ICR instrument from DHB. Replicate analysis gave coefficients of variation of less than 0.2. The standard protocol for N-glycan release from glycoproteins requires relatively long deglycosylation times (from several hours to, usually, overnight) and relatively high enzyme concentration (from 1:250 to 1:500 enzyme/substrate ratio). Szabo, Guttman, and Karger (2010a) have used a high-pressure method, both to reduce the reaction time and the amount of enzyme required. Thus, a pressure-cycling device was use to cycle the pressure from atmospheric to as high as 30 kpsi. Greater than 95% release of the Asn-linked glycans from bovine RNaseB, human TF, and polyclonal human immunoglobulin was achieved in only a few minutes using as low as 1:2,500 enzyme: substrate molar ratio. A reactor with immobilized PNGase F on a monolithic polymer support in a capillary has been developed that allows fast and efficient release of N-linked glycans. Performance was determined with RNaseB, chicken ovalbumin, and human IgG with detection by MALDI-TOF MS. The optimized reactor was integrated into a multidimensional system comprising on-line glycan release and hydrophilic interaction liquid chromatography (LC) followed by ESI-TOF MS detection. Using this system, human IgG was deglycosylated at room temperature in 5.5 min to an extent similar to that achieved with the soluble enzyme after 24 hr at 37˚C (Krenkova, Lacher, & Svec, 2009) . Immobilization of PNGase F on detonation nanodiamonds has resulted in glycan release from glycoproteins in less than 10 min . The method, using trypsin immobilization, also gave good results and proved to be better for proteolysis than the use of commercial immobilized trypsin beads. Artefacts associated with PNGase release of N-glycans. For solution release of glycans, the glycoprotein is usually denatured by reduction and alkylation or by use of detergents or other compounds such as urea. In general, low concentrations of urea (<3 M) do not usually cause irreversible protein denaturation. Indeed, PNGaseF itself is stable in 2.5 M urea at 37˚C for 24 hr and still possesses about 40% activity in 5 M urea. However, other glycoproteins appear more susceptible. For example, analysis by SDS-PAGE and MALDI-TOF MS have revealed that additional 2.5 kDa of glycans can be released by PNGase F if the deglycosylation is conducted in 2 M urea suggesting that urea treatment exposes a glycosylation site that was previously inaccessible to PNGaseF . Use of urea, however, can cause problems with the released glycans because it has been reported to compete with water for hydrolysis of the initially formed glycosylamines with the formation of a urea complex (Omtvedt et al., 2004) . Another artefact that has been found in glycans released with PNGase F involves the reaction of the glycosylamines with H 2 S to form the glycan-SH analogue. The H 2 S arises from dithiothreitol (DTT, 6/44), a reagent used for protein alkylation and present in some commercial preparations of the reagent. The consequence of this reaction is an increase in 16 Da giving the impression, from a simple mass measurement, that the glycan has an additional oxygen atom. Addition of 16 Da can also be observed in MALDI spectra as a consequence of the formation of [MþK] þ rather than [MþNa] þ ions, but this possibility can be excluded by formation of [MþCs] þ ions whereupon the 16 Da mass increase will still be present. The reaction with H 2 S was first noted with glycans from human IgG and negative ion MS/MS of the artefactual products located the 16 Da to the reducing-terminal GlcNAc residue. The negative ion fragmentation pattern was the same as that expected for a glycan with a hexose attached to the 6-position of the residue rather than fucose (deoxy-Hex) that was actually the case. This result emphasizes how careful one must be, not only in deducing compositions from glycan masses but also in interpreting their MS/MS spectra . Another artefact of the PNGase F release step, detectable by CE but not by MS has been identified as the product of epimerization of the terminal GlcNAc residue to N-acetylmannosamine (ManNAc, 62) under the slightly basic conditions usually employed in the release reaction. Reducing the pH to 5.5 effectively removed the by-product (Liu, Salas-Solano, & Gennaro, 2009h) . Other endoglycosidases. Another popular enzyme for N-glycan release is endo H which cleaves the chitobiose core of high-mannose and hybrid glycans but not complex ones, leaving a GlcNAc residue, with any linked fucose, attached to the protein. Pace et al. (2009) have made use of this property to release and identify minor glycans in IgG without interference from the more abundant complex glycans. Endo F1 has a similar specificity and has been used by, for example, Voutilainen et al. (2010) to detect glycosylation in Talaromyces emersonii cellobiohydrolase Cel7A produced in the yeast Saccharomyces cerevisiae. Ammonium hydroxide/carbonate-based chemical deglycosylation and PNGase A enzymatic release have been compared for glycan release from a plantibody produced in tobacco plants (Triguero et al., 2010) . Although both methods gave similar profiles as evaluated by HPLC of 2-AB derivatives, the main drawback of the chemical release method was that it induced degradation of a1,3-fucosylated N-glycans. ii. Extraction and purification of released glycans. Cleanup of samples prior to MS is crucial to obtaining good spectra. Many methods are in use; porous graphatized carbon (Buser et al., 2010) is popular and we have found Nafion membranes (Börnsen, Mohr, & Widmer, 1995) to be convenient at removing both salts and some hydrophobic compounds. Avoiding the introduction of contaminants is also important. Disposable plasticware such as plastic test tubes that are normally used to process samples have been shown to be a major source of contamination. The contaminants, which produce ions, mainly prompt fragments across the entire mass range to about m/z 3,000, originate from polymers that are used to protect the plastic against oxygen or UV light degradation. Such compounds are hindered amine light stabilizers (HALSs) used in modern polyolefin (polypropylene, polyethylene) stabilization. The polymeric agent: poly-(N-b-hydroxyethyl-2,2,6,6-tetramethyl-4-hydroxy-piperidinyl succinate, 63), known as Tinuvin-622 or Lowilite 62, has been found to leach from laboratory polypropylene or polyethylene plastic test tubes into solvents used for sample preparation. 1.5 mL plastic tubes were found to be the major source of the contamination but the authors of the paper found that this could be minimized by using large solvent volumes of, for example, matrix solution (Sachon et al., 2010) . Amano and Nishimura (2010) have used two kinds of hydrazide-functionalized glycobeads, termed GlycoBlot H and GlycoBlot ABC of which the latter carries an additional fluorescent probe, to extract PNGase F-released N-glycans from solution. Sialic acids were then converted into methyl esters using the 3-methyl-1-p-tolyltriazene (MTT, 6/23) reagent de-scribed by Miura et al. (2007) and the products were examined by MALDI-TOF MS after release of the glycans from the beads by mild acid hydrolysis. The method was used to examine human serum glycoproteins for cancer biomarkers. Full experimental details of the extraction and derivatization procedure are given in the paper. Enrichment of serum and cellular glycoproteins with Glycoblot H beads has also been used for O-glycan analysis . Glycans were released from human milk osteopontin and urinary MUC1 glycoproteins with ammonium carbamate and the method was proposed as ideal for identification of biomarkers. A method for sequentially enriching sulfated glycans by strong anion-exchange chromatography according to their degree of sulfation has been described by Lei, Novotny, and Mechref (2010) . The method is based on modifying the binding ability of strong anion-exchange material with different sodium acetate concentrations, thus enabling selective binding and a subsequent elution of different glycans according to their degree of sulfation. Before this enrichment, the negative charge on any sialic acid was eliminated by permethylation. The method was initially optimized using sulfated oligosaccharide standards and then used to examine the sulfated N-glycans from bTSH, a glycoprotein possessing mono-and disulfated N-glycans. f. Analysis of released glycans Ahn et al. (2010) have obtained good resolution of N-glycans as their 2-AB derivatives using HILIC columns packed with 1.7 mm sorbent. Glycans were released from RNase B with PNGase F and extracted using a microElution HILIC SPE 96well plate. The labeled glycans were also extracted from the preparative reagents using the same plate and their integrity was checked by MALDI-TOF MS. In another technique, Guillard et al. (2009) have experimented with optimizing a linear ion trap instrument for automated measurement of permethylated N-glycans in serum. Glycans were released with PNGase F, cleaned with graphitized carbon and permethylated with the sodium hydroxide method. DHB, although the favored matrix for carbohydrates, failed to give the necessary reproducibility because of the large crystal size. CHCA, on the other hand, proved to be satisfactory. Full experimental details for analysis of O-and N-linked glycans have been published by several authors (Azadi & Heiss, 2009; Morelle et al., 2009a; North et al., 2010b) . g. Total methods for glycoprotein structure There have been many reports of methods for total glycoprotein analysis of which the following are representative. A small-scale method for N-glycan release and analysis from plants used to produce recombinant glycoproteins has been described . Concentration, protease digestion and deglycosylation are carried out in a single concentrator unit Mass Spectrometry Reviews DOI 10.1002/mas without the need for intermittent purification. This approach minimized adsorptive losses and facilitated handling. The plant protein was concentrated in a unit with a 5 kDa cutoff and after buffer exchange, pepsin digestion was carried out in the concentrator overnight. Deglycosylation was carried out with PNGase A for 24 hr. Released N-glycans were purified using reversed-phase and cation exchange chromatography in micro-columns and analyzed by MALDI-TOF MS without derivatization. A chip-based reversed-phase LC/MS method for N-glycan analysis suitable for biomarker discovery has been developed by . N-Glycans were released from bovine fetuin as a model glycoprotein and human serum glycoproteins with PNGase F and reduced to alditols with an ammonia-borane complex. The glycans were then permethylated in dimethylformamide to avoid artefacts in MS measurements and their structures were checked by MALDI-TOF measurements. Reversed-phase microfluidic LC of the permethylated N-linked oligosaccharide alditols was then performed and was shown to resolve some closely related structures. Optimized LC gradients, together with nanospray MS were then used with human serum samples to distinguish breast cancer patients from control individuals. A previously established two-dimensional HPLC technique has been adapted as a HPLC-MALDI MS method for N-glycan analysis by Gillmeister et al. (2009) . Glycans were released from glycoproteins with PNGase F purified with graphitized carbon and fluorescently labeled with 2-AP. The labeled glycans were analyzed on a 2-mm reversed phase (RP) HPLC column and spotted onto a MALDI-TOF MS plate together with the DHB matrix using an automated plate spotter. The method gave a 100-fold reduction in the required amounts of starting protein compared with the earlier procedure. The entire process could be carried out in 2-3 days for a large number of samples as compared to 1-2 weeks per sample for previous two-dimensional HPLC methods. The modified method was verified by identifying N-glycans from an IgG antibody from human sera samples and applied to analysis of tissue plasminogen activator (TPA) from CHO cell cultures under varying culture conditions. Kim et al. (2009b) have released glycans on a polyvinylidine difluoride (PVDF) membrane with PNGase F and cleaned them with graphitized carbon contained in a 96-well plate before converting them into Girard's T derivatives to introduce a constitutive cationic charge for quantification. Analysis was by MALDI-TOF MS and the robust method was used to profile N-glycans from ovarian cancer patients using as little as 5 mL of serum. A high-throughput method for the analysis of human plasma glycomes using a 48-channel multiplexed capillary gel electrophoresis (CGE) DNA sequencer with laser-induced fluorescence detection (CGE-LIF) system has been described with MALDI-TOF MS used to provide structural information (Ruhaak et al., 2010c) . Glycans were released from plasma glycoproteins in a 96-well plate using PNGase F and converted into APTS derivatives with the help of 2-picoline borane (27) as the reducing agent. Analysis by CGE-LIF using the DNA sequencer allowed 96 samples to be analyzed in only 2.5 hr (the experimental time was longer because of two overnight incubations). The method was applied to a study of glycosylation patterns during first, second, and third trimesters of pregnancy, as well as 6 weeks, 3 months, and 6 months postpartum. Although analysis of glycoproteins carrying neutral glycans is now routine, analysis of glycoproteins with sialylated glycans is more difficult. Hao, Ren, and Xie (2010) have approached the problem by first performing a tryptic digestion to give peptides and glycopeptides. Peptide mass fingerprinting was performed on the peptides in order to identify the protein. The glycopeptides, separated by HILIC chromatography, were examined by MALDI-TOF MS and MS/MS and treated with PNGase F to release the glycans, which, together with the resulting peptides were again examined by MS. The Asn to Asp conversion in the peptide fraction enabled the glycosylation site to be identified. Finally, the glycans were desialylated with dilute HCl and again analyzed by MS. The technique was applied to glycoproteins from human serum separated by 2-D electrophoresis and the differences in N-glycosylation were successfully determined for a1-antitrypsin between different gel spots. In another method, sialylated glycoproteins have been selectively periodate-oxidized, captured on hydrazide beads, trypsinized and released by acid hydrolysis of the sialic acid glycosidic bonds. Mass spectrometric fragment analysis allowed identification of glycan structures and additional fragmentation of deglycosylated ions yielded peptide sequence information which allowed glycan attachment sits to be identified together with identification of the protein. Using this method, the investigators identified 36 N-linked and 44 O-linked glycosylation sites on glycoproteins from human cerebrospinal fluid . Related to this method is one developed by Klement et al. (2010) for enrichment of O-GlcNAc-modified proteins. Glycoproteins were again oxidized with periodate and captured by hydrazide resin capture. Rather than release of the peptide enzymatically, the glycopeptide was released by hydroxylamine treatment which also converted the aldehyde groups of the oxidized glycan to oximes. The open nature of the carbohydrate ring, following oxidation, lead to the production of characteristic fragment ions facilitating both glycopeptide identification and site attachment. The method was applied to analysis of a-crystallin A and the Drosophila proteaosome. h. Comparisons of methods for N-glycan analysis A comparative study of three techniques, MALDI-TOF, SDS-PAGE and CGE-on-a-chip, for measuring the MWs of large glycoproteins has been reported by Müller et al. (2010b) . It was found that all three techniques were capable of determining the MW of all high MW (glyco)proteins tested. The non-commercial CGE-on-a-chip assay allowed electrophoretic separation of proteins in the MW range from 14 kDa to 1 MDa. MW assignment was limited to 500 kDa in the case of SDS-PAGE but with the proper matrix (THAP for most glycoproteins, sinapinic acid for a2-macroglobulin) and sample preparation, analysis with a standard MALDI-TOF-MS provided accurate MWs for all high MW proteins up to 1 MDa. Three methods for N-glycan characterization, namely MALDI-MS of glycopeptides from tryptic digestion, negativeion ESI-MS/MS of released N-glycans, and normal-phase HPLC of fluorescently labeled glycans, in combination with exoglycosidase sequencing, have been evaluated for glycan identification using monoclonal antibodies expressed in tobacco plants as model compounds (Triguero et al., 2010) . The MS methods identified the major glycans, but the HPLC method was found to be the best for identification and relative quantitation. Negative-mode ESI-MS/MS easily provided direct identification of features such as the linkage position of the fucose residue linked to the inner core GlcNAc residue. Grey et al. (2009) have developed a high-performance ion exchange chromatographic method for N-glycan analysis and have shown that it gives very similar results to analysis by MALDI-TOF. A series of standard glycans was examined and the method was extended to the analysis of N-glycans released from IgG1. An inter-laboratory study involving eleven UK laboratories using their routine glycan analysis procedures looked at reproducibility on glycan profiling from N-glycans released before the study from four glycoproteins, human and bovine AGP, bovine pancreatic RNaseB and human serum immunoglobulin G (hIgG). Data interpretation focused on the relative amounts of different glycan structures present, the degree of sialylation, galactosylation profiles, fucosylation, and bisecting GlcNAc content and the number of glycan components identified. All laboratories found high levels of sialylation for human and bovine AGP, but varying amounts of di-, tri-, and tetra-antennary glycans. Values obtained from mass spectrometric and chromatographic methods clustered separately. The proportion of the major Man 5 GlcNAc 2 from RNaseB was between 29% and 62%. Proportions of fucosylated and bisected GlcNAc glycans from hIgG were between 58% and 96% and 9% and 23%, respectively. Mass spectrometric approaches consistently identified more glycan species, especially when both N-glycoylneuraminic acid (Neu5Gc) and Neu5Ac were present (Thobhani et al., 2009) . A recent test of the ability of several laboratories to identify N-glycans released with PNGase F from a mixture of four glycoproteins, asialo-fetuin, chicken ovalbumin and both human and bovine AGP has also yielded some alarming results (Orlando, Leymarie, & Keck, 2010) . Although 18 of the 19 laboratories detected the presence of fucosylated complex Nglycans, 14 of them incorrectly located the fucose to the core GlcNAc of human AGP rather than to an antenna. All nine of the labs using MS (not MS 2 ) misidentified the site and all five of laboratories relying on software to identify the site also reported it incorrectly. Although the ionic charge was not specified, it is assumed to be positive because it would be impossible to make this mistake with negative ion fragmentation. Features such as the position of fucose residues produce diagnostic cross-ring fragments whose mass depends on the location of the fucose residue . Such mis-identifications are particularly worrying because serum AGP is elevated in inflammatory disease and is of potential use as a biomarker. Many laboratories using this potential biomarker in serum also report the structure incorrectly. Another problem in the survey arose with N-glycoylneuraminic acid (5/38), present in the bovine version of a1glycoprotein. This carbohydrate is antigenic and is of concern to pharmaceutical companies producing pharmaceuticals in organisms that utilize this sialic acid. In the survey, eight of the laboratories, including seven of the eight participating industrial laboratories failed to detect its presence. All of the four laboratories that used a fluorescence tag, failed to detect this sialic acid. Slightly better results were obtained by laboratories using MALDI; of ten labs that used this technique, only two failed to detect this sialic acid. Most laboratories that successfully detected this carbohydrate, permethylated their samples, which, of course, would stabilize it to MALDI conditions. With mixtures containing different quantities of glycans, no lab correctly detected either three or four of the changes, one lab identified two of the four changes, seven labs identified one change but the 11 other labs failed to identify any changes correctly. Clearly, current analytical methods leave much to be desired. i. Applications of MALDI to the detailed structural determination of N-linked glycans A large number of reports have appeared on the applications of the above techniques to analysis of N-glycans from specific glycoproteins. These are summarized in Tables 10 and 11 . Other examples can be found in the tables on medical applications of MALDI MS (Table 23 ) and biopharmaceuticals (Table 24) . Some of the more unusual structures to be discovered are tetraantennary glycans with poly-N-lactosamine extensions with up to nine fucose residues in human seminal plasma , a Man 5 GlcNAc 2 glycan with a bisecting GlcNAc residue (64) (Buser et al., 2010) ; a Man 8 GlcNAc 2 -type glycan with two bisecting GlcNAcs (65) proposed from Dictyostelium discoideum Feasley et al., 2010) and an unusual glycan with internal fucose and glucuronic acid (GlcA) from Rapana venosa hemocyanin (Velkova et al., 2009) . However, in the latter case, the structure was based on evidence from only one positive ion MS/MS spectrum and is open to alternative interpretations. Symbols as defined for structures 17 and 18 plus = mannose j. Miscellaneous studies Among other studies, MALDI-TOF MS has been used to confirm the glycan compositions of several well-known glycoproteins in a study showing that glycosylation protects proteins against free radicals generated from toxic xenobiotics (Martínek et al., 2010) The rice-derived recombinant human transferrin (rhTF) has been shown to be non-N-glycosylated by MALDI and PNGase F enzyme digestion . Structural determination (highmannose, with bisect on branching mannose and 6antenna) (Continued) (Continued) sodium hydroxide followed by reduction to prevent a peeling reaction is the most common method but has the disadvantage of eliminating the reducing terminus, thus preventing tagging with fluorescent or other tags. Some investigators have, thus, investigated the use of milder bases with the aim of avoiding the reductive stage and retaining the reducing terminus. Zheng, Guo, and Cai (2009) have compared ammonia, methylamine and dimethylamine at 55˚C for 6 hr for release of N-acetylgalactosamine (GalNAc) from a small glycopeptide. b-Elimination with dimethylamine and methylamine resulted in the conversion of the glycopeptide to 69.2% of the dimethylamine derivative at m/z 550.32 and 61.5% of the methylamine derivative at m/z 543.33, respectively. However, the incubation of the glycopeptide with ammonia only resulted in 8% production of the product. The authors concluded that elimination with dimethylamine was the most efficient for release the O-linked glycans. Release with methylamine was used by Sun et al. (2010b) to determine the glycosylation sites in human protein C inhibitor by the 13.03 mass increment introduced by the reaction. Release with ammonia has been investigated in detail by Yu et al. (2010a) for O-glycan chains with b1,3-linked cores. In contrast to b1,4-linkages of the N-glycan-type, which were shown to be stable under the ammonium-based alkaline conditions, the b1,3-linkage was found to be labile and to give considerable peeling . The results indicated that complete prevention of peeling under nonreducing alkalicatalyzed hydrolysis conditions remains difficult. The yields of O-and N-glycans from bovine fetuin released by conventional means (PNGase F and reductive b-elimination with NaOH) were found to be greater. It was concluded that great care should be taken when employing such non-reducing alkaline conditions in glycomic analysis and in obtaining some O-glycans for functional studies. Because the hydroxide ion appears to cause the unfavorable peeling reactions, Miura et al. (2010b) have investigated the use of the ammonium salt, ammonium carbamate for glycan release. The efficiency of release with ammonium carbamate was compared with a common conventional procedure, namely saturated ammonium carbonate/aqueous ammonia with bovine submaxillary mucin (BSM) as the test compound. Release with ammonium carbamate did not exhibit significant loss of GlcNAc-b1,3 (Neu5Aca2,6)GalNAc or GlcNAcb1,3(Neu5Gca2,6)GalNAc Structural determination (highmannose, bisected high-mannose glycans). Development associated with alteration of fucosylated glycans Structural determination (highmannose, hybrid, complex glycans). Poly-fucosylation Human (HT-29 5M12 colon cancer cells) PNGase F, TOF, glycans (per-Me) Pattern of N-glycosylation serves as a recognition signal for galectin-4. High-Man, hybrid, bi-, tri-tetra-antennary. (Stechly et al., 2009) Human (Cytolytic T lymphocytes) PNGase F, TOF/TOF (DHB), glycans (per-Me) Loss of effector function shown to be accompanied by major changes in Nand O-glycosylation (Antonopoulos et al., 2012) Marmota monax (woodchuck with liver cancer), Gal-α-(1→3)-Gal in mouse epidermis but GalNAc-β-(1→4)-GlcNAc in human. High-mannose, hybrid complex (Uematsu et al., 2009) PNGase A PNGase F, TOF/TOF (DHB/3% Plants engineered to produce Lewis X epitopes. (Paucimannosidic glycans) Mass Spectrometry Reviews DOI 10.1002/mas and the profile of the major O-glycans was similar to that obtained following conventional reductive amination with NaOH/NaBH 4 . On the other hand, glycans obtained by treating BSM with ammonium carbonate/28% aqueous ammonia and analyzed by MALDI-TOF MS showed a significant increase of the disaccharide components, Neu5Aca2,6GalNAc and Neu5-Gca2,6GalNAc, suggesting the presence of a peeling reaction. Use of ammonium carbamate, thus, appears to produce efficient release without concomitant peeling. The release was performed by addition of powdered ammonium carbamate and incubation for 20 hr at 60˚C. Yamada et al. (2009) have used a recently developed automated release apparatus using lithium hydroxide to obtain O-glycans from leukemia and epithelial cancer cells. Because these cells usually contain free glycans, the investigators first reduced these with sodium borohydride and then labeled the released glycans with 2-AA in order to avoid interference. A new release method reported by Goetz, Novotny, and Mechref (2009a) combined enzymatic and chemical techniques and used b-elimination to cleave glycans from serine (Ser) and threonine (Thr) but not Asn. The method involved first a nonspecific proteolysis with pronase, followed by solid-phase Hydrazine, TOF (DHB), glycans (2-AP) Structural determination. High-Man, hybrid, bi-, tri-, tetra-antennary complex (Sumiyoshi et al., 2010) Rabbit and chicken (Erythrocytes) TOF, glycans (2-AP) To study alterations in receptor-binding properties of H1-type swine influenza viruses in embryonated chicken eggs. High-Man, hybrid, bi-, tri-antennary (Takemae et al., 2010) Rat ( Structural determination (highmannose, hybrid, complex glycans) Girard's T for quantitation (Kim, et al., 2009c) Sus domestica (Respiratory epithelial cells) PNGase F, TOF, TOF/TOF (DHB), glycans (per-Me) Influenza virus shown to utilise α2→6linked Neu5Ac to infect cells permethylation with sodium hydroxide. The basic sodium hydroxide caused the glycans to be released by b-elimination and these were immediately permethylated. This combination of the enzymatic and chemical procedures was reported to give a substantial improvement in sensitivity and analytical reproducibility over existing methods by minimizing sample losses. Moreover, the approach was reported to extend the cleavage protocols to large glycoproteins where small oligosaccharides were not accessible to conventional chemical treatment. The method was developed with fetuin and used to identify new O-glycans from bile salt-stimulated lipase (BSSL). Maniatis, Zhou, and Reinhold (2010) have released O-glycans with aqueous dimethylamine in the presence of sodium borohydride by use of a microwave oven. The release was performed at 70˚C and, for a heptapeptide carrying a GlcNAc group attached to Thr, was complete in 70 min. The reaction also labeled the site of detachment with a dimethylamino group. Use of a 1:1 mixture of dimethylamine and [ 2 H 3 ] 2 NH produced doublets in the peptide mass spectrum separated by six units, allowing the glycosylation sites to be readily identified. N-Glycans are frequently removed from glycoproteins before O-glycan removal by b-elimination. However Stone et al. (2009) have reported improved recovery of O-glycans from murine tissues without prior N-glycan removal. KBH 4 and KOH were used to remove the O-glycans and possible low levels of concomitantly released N-glycans were tolerated. ii. Use of hydrazine. Hydrazine has also been used to release these glycans with a new gas-phase method using anhydrous hydrazine being evaluated by Goso, Tsubokawa, and Ishihara (2009) with MUC-type oligosaccharides from porcine gastric mucin (PGM) and bovine fetuin. Released glycans were examined by HPLC and MALDI-TOF as 2-AA derivatives. Glycans obtained by the treatment with hydrazine at 65˚C for 6 hr resembled those obtained by b-elimination, except for the additional disaccharide fractions derived from the core 1 side of the oligosaccharides by further degradation. The other degraded products derived from the core 2 side could not be derivatized by 2-AA, therefore, were not visible by fluorescence detection. Release of the glycans was incomplete after 6 hr but almost complete liberation was achieved by extending the treatment to 18 hr. However, degradation increased. In this case, the addition of barium oxide to the reaction vessel decreased the degree of further degradation. Results similar to PGM were obtained from bovine fetuin, but with less degradation. Application of this method to the analysis of rat gastric mucin (RGM) showed that RGM has a large oligosaccharide portion on the core 1 side. iii. Other methods. A new method for O-glycan removal for study of the residual deglycosylated protein has been reported . Desialylated glycoproteins whose sugar chains consisted of Gal-GalNAc, were immobilized on alkali-stable, reversed-phase Poros 20 beads and treated with periodate to oxidize the cis-glycol groups in the Gal residue. The resulting aldehydes were then susceptible to b-elimination under mild (NH 3 ) basic conditions. The remaining GalNAc residue, which now contained a cis-glycol group, was oxidized with further treatment with periodate and removed with base. Although the number of cycles required depended on the number of Gal-GalNAc repeats, large core 2-type glycans that usually only have Gal attached to the 3-position of the core GalNAc, could be deglycosylated in only two steps. O-Linked glycosylation often occurs in MUC-type domains that are heavily and heterogeneously glycosylated. Several strategies to determine the heterogeneity of these domains have recently been investigated with four glucanases secreted in large quantities from Trichoderma reesei, all of which contained heavily O-glycosylated MUC-like linker regions, being used as models. The strategies involved monosaccharide compositional analysis and identification of the released glycans by HPAECpulsed amperometric detection (PAD) and carbon-LC ESI-MS/ MS. Glycosylated peptides were generated by different protease digestions (trypsin, papain, Asp-N, PreTAQ) and enriched by HILIC microcolumns. The complex O-glycan heterogeneity was determined by MALDI-MS and ESI-MS, but the dense O-glycosylation in the MUC-type domains conferred high resistance to protease cleavage. ETD-MS/MS of the glycopeptide-enriched protease digests was unsuccessful for the assignment of O-glycosylation at individual sites within the MUCtype domains but allowed several previously unknown O-linked sites outside the defined linker region to be found on two of the four glucanases (Christiansen et al., 2010) . iv. Comparison of methods. Three samples of IgA1 isolated from the serum of patients with multiple myeloma have been distributed on behalf of the Human Proteome Organization Human Disease Glycomics/Proteome Initiative to 15 laboratories for comparative analysis of their O-glycans. A range of techniques was used; the two strategies that yielded the best data were direct positive ion MS analysis of permethylated glycans and LC-MS analysis of native reduced glycans in negative ion mode. The studies reinforced the pre-eminent performance of MS techniques for O-glycan profiling (Wada et al., 2010b) . Several reviews and general analytical methods have been reported; these are listed in Table 14 . Loss of sulfate is a major problem in the MALDI analysis of these compounds but this can be avoided by use of desorption ESI (Przybylski et al., 2010a) . Little use appears to have been made of the peptide binding method for stabilization of sulfate groups in these compounds as reported by Juhasz and Biemann (1994) . One report concerns the identification of a pentasulfated hexasaccharide responsible for binding to the growth factor pleiotrophin . The compound was complexed with (Arg-Gly) 15 and identified by MALDI-TOF from DHB. In another study , a pentasulfated hexasaccharide with a novel structure (D 4,5 HexAa1-3GalNAc(4S)b1-4IdoA(2-S)a1-3GalNAc(4S)b1-4IdoA(2S)a1-3GalNAc(4S)) has been isolated from the chondroitinase AC-I digest of shark skin. Again, (Arg-Gly) 15 was used as the complexing agent. Bultel et al. (2010) have developed a method for analysis of heparin oligosaccharides by using controlled nitrous acid degradation followed by high-performance anion exchange chromatography (HPAEC) separation and UV-MALDI-TOF analysis. The use of three different matrices, DHB, CHCA, and nor-harmane were investigated but only DHB and nor-harmane were needed to assign the position of sulfate groups. DHB allowed the molecular ion to be detected in nearly all cases and gave fragments arising from the loss of sulfate groups. Nor-harmane, in contrast, produced mainly fragments. In all cases, ions retaining the sulfate groups were observed making these fragments essential for assigning the sulfated positions of each residue. While nor-harmane was not able to produce enough analyte desorption/ionization, fragments useful for structural assignment were produced by the addition of butylammonium formate to the DHB matrix. A method for analysis of GPI anchors on the "proteomic" scale has been described . Partially purified proteins were separated by SDS-PAGE and then blotted onto a PVDF membrane. Following identification of the protein, the GPI anchor was analyzed by three methods. First, the compound was hydrolyzed with HCl in the presence of [1,2,3,4,5,6-2 H 6 ]myo-inositol and the hydrolysate was analyzed as trimethylsilyl (TMS) derivatives by GC/MS. Next, the phosphate bonds were cleaved and the carbohydrate structure was elucidated by electrospray or MALDI-TOF MS. Finally, the diacylglycerol-attached myo-inositol moiety was detached by nitrous acid deamination and analyzed by negative ion electrospray. Bütikofer et al. (2010) have studied lipid remodeling of GPI glycoconjugates in procyclic-form trypanosomes and shown that, in Trypanosoma congolense, the steady-state lipids consist of lyso-(acyl)phosphatidylinositol (PI, 66), deacyl-PI and deacyl-(acyl)PI species, where (acyl) indicates an acyl group attached to the inositol moiety. MALDI-QIT-TOF in negative ion mode from THAP was used to analyze the PI species after deamination with nitrous acid. Structural determination (Core 1, core 2). Samples from geographically remote labs. Similar glycans (Babu, et al., 2009) Human, (enterocytelike HT-29 cells) β-Elimination, TOF, glycans (per-Me), GC/MS Structural determination (Core 1) (Morelle, et al., 2009b) Human ( Absence of fucose on Core 3 glycans impairs BabA-mediated adhesion to gastric mucosa (Magalhães et al., 2009) (Continued) Although many bacteria produce S-layer proteins with glycosylation, Qazi et al. (2009) have used MALDI-TOF MS to show that those from Clostridium difficile are not glycosylated. This topic has been reviewed by Zhang et al. (2009f) and Capote and Sanchez (2009) . MALDI is used mainly to determine the extent of glycation of intact proteins or to determine the sites of attachment of the covalent glycans. a. Specific methods for glycated peptides Amadori peptides have been enriched with boronate affinity tips for measurement by MALDI-TOF/MS. The tips showed the highest binding efficiency for glucose at pH 8.2 employing ammonium chloride/ammonia buffer with ionic strength of 150 mM. The bound constituents were released by sorbitol (1/ 42) or formic acid. Using sorbitol for elution required desalting prior to analysis. Of three different sorbents tested: fullerenederivatized silica, ZipTips (C18), and C18 silica, fullerenederivatized silica and ZipTips showed the same performance with respect to the numbers of glycated peptides and gave better performance than C18 silica. Fewer glycated peptides were detected by LC-MS/MS than by MALDI . A novel fullerene(C60)-derivatized silica material has been compared with octadecyl(C18) and triaconthyl(C30)-silicas for their ability to recover peptides from digests of HSA and fibrinogen. C30-and particularly the C60(30 nm)-SPE materials were found to be the two most effective. After glycation the digests of fibrinogen and HSA were also separated. This new method made the detection of a considerably higher number of glycated peptides possible compared to the unfractionated digests and the use of boronate affinity chromatography in the case of fibrinogen. For HSA, 10 new sites of glycation at Lys and Arg residues were found . A mass spectrometric method for screening large tandem mass spectrometric (MS/MS) datasets for protein glycation with glucose (1/4), lactose (67) and maltose (68) has been developed (Montgomery, Tanaka, & Belgacem, 2010) . Control experiments using a standard peptide containing a single glycation site led to the discovery of characteristic neutral loss fragmentation patterns in MS/MS analysis for glucose, lactose and maltose condensed with peptide. For glucose glycation, neutral losses of 36, 120, and 162 Da were observed in accordance with previously published reports. The neutral loss patterns for lactose and maltose were found to be identical, with characteristic losses of 162, 198, 282, and 324 Da. These signature losses were observed irrespective of the MALDI mass spectrometer used and were valid in both TOF-TOF and QIT-TOF instruments. These neutral loss signatures were then applied to elucidation of modified peptides from a complex HSA digest glycated with each of the proposed sugars. Screening of these large datasets O-labeled digests indicated that Lyss 525 and 439 also had significant degrees of modification. The modifications that occurred at these sites were variations of fructosyl-Lys and advanced glycation end products (AGEs) which included 1-alkyl-2formyl-3,4-glycoyl-pyrole (4/45) and pyrraline (4/44). The fragmentation behavior of the peptide Ac-PAAPAA-PAPAEKTPV-OH (human histone H1.4, positions 6-20) glycated via its Lys residues to ADP-ribose has been studied by Fedorova, Frolov, and Hoffmann (2010a) . Under MALDI conditions, the ADP-ribosyl group was cleaved, almost completely at the pyrophosphate bond by ISD and PSD. However, this cleavage was very weak in ESI-MS. The remaining phospho-ribosyl group was stable, providing a direct and reliable identification of the glycation site via the b-and yion series. As well as being associated with health problems, in for example, diabetes, protein glycation is important in the food industry in, for example, the browning of food during cooking. The reaction is also being used to attach carbohydrates to proteins to improve technological and biological functionalities. In relation to this latter use, Corzo-Martnez et al. (2010) have used MALDI-TOF MS to study the reaction between b-lactoglobulin and the sugars galactose and tagatose (69) and found that the reaction can be competitively moderated with pyridoxamine (70). Other reports of the use of MALDI to study glycation of specific proteins are summarized in Table 15 . Typical structures consist of GlcNAc-MurNAc (71) disaccharides cross-linked by short peptides. They are usually analyzed as muropeptides following enzymatic digestion. Reports of the use of MALDI to study peptidoglycans and muropeptides are summarized in Table 16 . This is a very large group of compounds but most work involving MALDI has been concentrated on the LPS from bacteria and the GSLs. Work, mainly involving structural determination, on the many other types of glycolipids found in bacteria and similar organisms, is summarized in Table 17 . A general review on analysis of microbial glycopolymers has been published by , and Fuchs and Schiller (2009) phenol/chloroform/petroleum ether and the molecules are frequently split into their component parts by mild acid hydrolysis for further structural analysis. Dephosphorylation and deacylation are also common. Reviews on the structural investigation of bacterial LPS by MS and MS/MS have been published by Banoub et al. (2010) and by Grice and Wilson (2009) . Lipid A from Coxiella burnetii, the causative agent of Q fever has been discussed by Toman, Skultety, and Ihnatko (2009) and a more general review of the core region and lipid A of LPS has been published by Holst and Molinaro (2010) . The first structures of LPS to be elucidated from cyanobacteria has been reported by Snyder et al. (2009) . Two strains of marine Water-soluble proteins Glucose L-TOF (2,6-DHAP) Use of MALDI-TOF to study glycation during malting Synechococcus, WH8102 and CC9311, were used and were shown to have very simple structures without the complex O-chain found in most proteobacteria. The LPS (72) of these cyanobacteria did not contain phosphate, heptose (Hep) or Kdo (1/13) but instead possessed 4-linked glucose as their main saccharide component, with low levels of GlcN and galacturonic acid (GalA). MALDI-TOF MS of the intact minimal core LPS revealed triacylated and tetraacylated structures with a heterogeneous mixture of both hydroxylated and nonhydroxylated fatty acids connected to the di-GlcN backbone. In contrast to enteric lipid A, which can be liberated from LPS by mild acid hydrolysis, lipid A from these organisms could be produced only by two novel procedures: triethylamine-assisted periodate oxidation and acetolysis. Unique caryophyllose (a-3,6-dideoxy-4-C-(D-altro-1,3,4,5tetrahydroxyhexyl)-D-xylo-hexopyranose, 73)-containing cell wall glycolipids have been identified in LPS from Mycobacterium marinum (Rombouts et al., 2009) . MALDI-TOF spectra were obtained from DHB and ESI-MS/MS was used to elucidate the glycan sequence. b. Lipid A Sample preparation has been shown to be critical for assessing the true composition of these compounds. In a comparative study, Kawasaki (2009) have shown that MALDI-TOF MS analysis of lipid A prepared using a commercial "Tri-reagent"based procedure with a 5-chloro-2-mercaptobenzothiazole (CMBT) (1/33) matrix gave the best results for compounds with a phosphoethanolamine (PEtN) modification. In contrast, the analysis of lipid A prepared using an LPS extraction kit-based procedure with DHB was preferable for the detection of an aminoarabinose modification. For isolation of lipid A from Yersinia enterocolitica, Pérez-Gutiérrez et al. (2010) used an ammonium hydroxide-isobutyric acid method. Lyophilized crude cells were incubated with isobutyric acid and ammonium hydroxide (5:3, v/v) at 100˚C for 2 hr, washed twice with methanol and the insoluble lipid A was solubilized in chloroform-methanol-water (3:1.5:0.25, v/v/v). Analysis was by MALDI-TOF MS from DHB in negative ion mode because of the presence of two phosphate groups. The lipid as A contained both four or six fatty acyl chains whose (Boniface et al., 2009) ratio changed with growth temperature. A similar method has been used by March et al. (2010) to study lipid A from Acinetobacter baumannii. The molecules were found to have from four to seven acyl groups. A microwave-assisted method for obtaining Lipid A from Helicobacter pylori has been developed by Zhou et al. (2009a) . Lyophilized cells were suspended in sodium acetate buffer (pH 4.5) containing proteinase K and subjected to microwave irritation at 50 W for 5 min at 58˚C and then kept for 1 hr at 100C . After centrifugation and washing, the dried supernatant was examined by MALDI-TOF/TOF from CMBT. The reliability of the technique was demonstrated by analysis of the lipid A from bacterial cells of different H. pylori strains. The phosphorylation and acylation patterns could be elucidated using material from a single colony. Furthermore, the investigators found unusual heptaacyl lipid A species present in low abundance in H. pylori mutant that have not been previously reported. The study was claimed to provide the first characterization by MS of the lipid A component from a single bacterial colony. The mass spectrometric behavior of lipid A is highly dependent on both the matrix and phosphorylation patterns. Zhou et al. (2010a) have investigated the effects of different matrices and co-matrices using lipid A from Escherichia coli O116 as a model system. Good results were obtained with CMBT with added EDTA (5/43) ammonium salt as the matrix. This matrix system was found to enhance the sensitivity of the detection of diphosphorylated lipid A by more than 100-fold and, in addition, provided tolerance to high concentrations of SDS and to both sodium and calcium chlorides at mM concentrations. The method was evaluated for analysis of lipid A with different phosphorylation patterns and from different bacteria, including H. pylori, Salmonella enterica serovar Riogrande, and Francisella novicida. An LC/MS-based assay has been developed for the quantitation of aminosugars, including GlcN (74), galactosamine (GalN, 75) and aminoarabinose (AraN, 76) together with ethanolamine (EtN), present in lipid A and has been applied to the analysis of lipid A isolated from several biosynthetic and regulatory mutants of S. enterica serovar Typhimurium and Francisella tularensis subspecies novicida characterized by MALDI-TOF. Lipid A was treated with TFA to liberate and deacetylate individual aminosugars and mass tagged with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (77), which reacts with primary and secondary amines. The derivatives were separated using RP-chromatography and analyzed with a quadrupole MS to detect quantities as small as 20 fmol. GalN was detected only in Francisella and AraN only in Salmonella, while GlcN was detected in lipid A samples from both species (Kalhorn et al., 2009) Use of grazing-incidence X-ray scattering and Monte Carlo simulation to investigate physics of bacterial survival against cationic antimicrobial peptides . (Continued) c. Core oligosaccharide During an analysis of the permethylated derivative of the core oligosaccharide from Aeromonas hydrophila, prepared by the Hakamori method, ions appearing at 78 mass units higher than the [MþH] þ ions were observed. These ions were determined to be dimethylsulfoxide (DMSO) covalent addition products resulting from the Michael addition of the dimsyl anion on the C-2-C-3 double bond of a 4,8-anhydro Kdo (1/13) residue followed by an addition of a proton on the double bond. Corresponding ions were also seen in methylations performed using the NaOH technique and represent the first characterization of these addition products (Sioud et al., 2010) . Typical pre-MALDI techniques for the analysis of these compounds include separation by TLC and cleavage of the Cer portion so that the glycan can be analyzed without the heterogeniety produced by the lipid. Reviews on the analysis of GSLs are summarised in Table 18 . a. Analysis of intact compounds Stübiger et al. (2009) have separated lipids, including GSLs by high-performance TLC, stained them with Coomassie blue and analyzed them either directly from the TLC plates or, after their removal, with an appropriate solvent. THAP (1/44) was used as the matrix with acetone as the solvent for the on-plate analysis because its high volatility minimized sample spreading. A method for structural profiling of individual GSLs in a single thin-layer chromatogram by multiple sequential immunodetection has been developed by Souady et al. (2009) . Structures of the antibody-detected GSLs were determined by direct coupling of TLC with IR-MALDI after treatment of the TLC plate with glycerol. This combined technique was used to demonstrate structural GSL profiling of crude lipid extracts from human hepatocellular cancer. A new method for analysis of GSLs involves selective ozonolysis of the C-C double bond in the ceramide moiety of biological samples and subsequent enrichment of the generated GSL aldehydes by chemical ligation using aminooxy-functionalized gold nanoparticles (aoGNP). The GSL-bound nanoparticles were removed by ultrafiltration and the GSLs were analyzed by MALDI-TOF and -TOF/TOF MS from DHB. The method was used for structural profiling of mouse brain gangliosides such as GM1, GD1a/GD1b, and GT1b for adult or GD3 in the case for the embryonic mouse. Because the saturated acyl groups remained intact, the spectra provided information on both the carbohydrate and fatty acyl moieties (Nagahori, Abe, & Nishimura, 2009) . The direct structural characterization of microbial GSL receptors by use of the TLC overlay assay combined with IR-MALDI-o-TOF-MS has been described (Müsken et al., 2010) . Glycan mixtures were separated by TLC in three parallel lanes. One lane was stained with orcinol and a second was overlayed with GSL-specific bacteria. The bound microbes were detected with primary antibodies against bacterial surface proteins and the relevant GSLs were detected in the third lane by IR-MALDI-TOF. The combined method worked on the microgram scale of GSL mixtures and was successfully applied to the compositional analysis of globo-series neutral GSLs recognized by P-fimbriated E. coli bacteria, used as model microorganisms for infection of the human urinary tract. (Niedziela et al., 2010b) Incubation of botulinum neurotoxin serotype D with the GSL, GT1b has produced a complex (MW 51,921) that was detected intact by MALDI-TOF MS from sinapinic acid and provided evidence that the toxin attacks neurons in a ganglioside-dependent manner (Strotmeier et al., 2010) . A method for generation of novel fluorocarbon derivatives from GSLs has been described by Li et al. (2010f) . The derivatives had high affinity for fluorocarbon phases allowing them to be recovered from biological matrices by fluorous solid phase extraction (F-SPE). Sphingolipid ceramide N-deacylase was used to remove the fatty acid from the ceramide moiety, after which the fluorocarbon-rich substituent (F-Tag, 78) was linked to the free amine. Finally, the molecules were permethylated for MS analysis and the method was used to examine a crude ganglioside mixture extracted from bovine brain. In addition, the flourous tag was used in a microarray format to fix F-tagged GM1 ganglioside to a fluorous glass surface, with the glycan intact and available for interaction with a fluorescent derivative of cholera toxin B chain. Cheng et al. (2010a) have used 9-AA (6/18) as a matrix for quantitative analysis of sulfatides in biological samples. The matrix was said to promote selective ionization of sulfatides in negative ion mode with a detection limit in the high attomole range. Experimental details have been published for highperformance TLC separation of glycolipids followed by blotting to a PVDF membrane in a technique termed Far-Eastern blot with analysis by MALDI-TOF MS (Taki et al., 2009) . b. Studies on the glycan moiety following removal of the ceramide For studies of the carbohydrate portion of these molecules, lipid heterogeniety is frequently reduced by removing the ceramide with enzymes such as Rhodococcal endoglycoceramidase or leech ceramide glycanase. Li et al. (2009e) have described the preparation of the intact oligosaccharides from GM1 (NeuAcGgOse 4 Cer) and GbOse 4 Cer as examples to show the use of ceramide glycanase and have optimized the specificity and detergent requirements of Rhodococcal endoglycoceramidase for the release of glycan chains from various GSLs. A novel method of detecting 6-gala series GSLs (those possessing an R-Galb1-6Galb1-1-Cer, group) has been reported (Ishibashi et al., 2009) . The method used the specificity of endogalactosylceramidase, an enzyme that is capable of hydrolyzing 6-gala series GSLs to produce intact oligosaccharides and ceramides but which also catalyzes transglycosylation reactions. In the latter reaction, the enzyme transferred oligosaccharides from the GSLs to acceptors such as fluorescent 1-alkanols. In this application, 7-nitro-2,1,3-benzoxadiazole pentanol (NBD-pentanol, 79) was used as an acceptor. The fluorescent products, NBD-pentanol-conjugated-6-gala oligosaccharides, were separated and detected by TLC or HPLC with a fluorescent detector and characterized by MALDI-TOF MS. The method could also be applied to glycoglycerolipids and digalactosyldiacylglycerol and was successfully applied to detect 6-gala series GSLs in the fungus, Rhizopus oryzae and the parasite, Taenia crassiceps. Other applications of MALDI to the analysis of these compounds is summarised in Table 19 . Rhamnolipids are glycolipids of the type 80 produced by Pseudomonas spp. MALDI-TOF MS approaches have been developed for high-throughput screening of naturally occurring (fungus), mycelia TOF (CHCA), PSD Ident. of novel neogala-series glycosphingolipids with terminal Man and Glc (Tani et al., 2010) Human (mesenchymal stem cells from bone marrow) Macrobdella decora endoglyco-ceramidase, R-TOF/TOF, FT-ICR (DHB), glycans (per-Me) Structural determination (Heiskanen, et al., 2009) Human (colonic adenocarcinoma cell lines) Ceramide glycanase, TOF/TOF, Q-TOF (DHB), glycans (per-Me) Enhanced expression of β3-Gal-T 5 activity induces in vivo synthesis of extended type 1 chains on lactosyl-Cer Human (umbilical vein endothelial cells) TOF (CHCA) Acyl mainly 24:0. May act as biomarker for inflammation, GSL = Gb4 (Okuda et al., 2010) Human (umbilical vein endothelial cells) TOF (CHCA) Structural determination and dynamics of globotetraosylceramide under TNF-α stimulation (Okuda, et al., 2010) Human (kidney and colon) TOF Identification of GSLs that bind shiga toxin from E. coli Human (embryonic stem cells) Switching of the core structures of GSLs from globo-and lacto-to ganglio-series on cell differentiation Miniature pig (endothelial and islet cells) Ceramide glycanase, R-TOF (DHB), ESI (-ve), glycans (per-Me and Girard's T) Structural determination. Identification of Neu5Gc epitopes ) (Krambeck, et al., 2009) Mouse (thymus) TOF/TOF (DHB), ESI Structural determination and natural killer T cell development (Li et al., 2009f) Mouse (liver and serum) TOF (-ve) Acute kidney injury down-regulates gene expression of hepatic cerebroside sulfotransferase. Quant. by MALDI-TOF. (Zhang et al., 2009h) Mouse ( Use of 9-aminoacridine (9-AA) as matrix, Structural determination -phospholipids, Gal-Cer-sulfate, Gb5 Trichoderma viride (fungus) L-TOF (7-NH 2 -2-Mecoumarin) Ident. of phosphocholine-containing glycosyl inositol phosphoceramides respectively. The method was validated by compositional analysis using GC/MS, fractionation by RP-HPLC and analysis by 1 and 2D NMR Applications of MALDI to the analysis of other glycolipids are listed in Table 20 . Although much work has been published on glycosides during the review period, MALDI appears to occupy a relatively minor position with fast atom bombardment (FAB) and, particularly ESI being the preferred techniques. Most work has been on the identification of glycosides from various plant sources using a variety of techniques such as nuclear magnetic resonance (NMR), UV and IR spectrometry. Applications of MALDI to the analysis of glycosides and other natural products are summarised in Table 21 . Several investigators have reported that the main fragmentation pathways of flavonoids are apparently independent of the ionization mode (ESI, atmospheric pressure chemical ionization (APCI), or MALDI) or the types of analyzers used to acquire the spectra (triQ, IT, or QTOF) as reported in a review of the structural characterization of flavonoid glycosides by multistage MS (Vukics and Guttman, 2010) . MALDI-TOF (DHB) was used by Bankefors, Nord, and Kenne (2010) to examine saponins from Quillaja saponaria bark extracts in connection with the development of a multidimensional method using HPLC and ESI-IT MS n for profiling complex mixtures of natural products. Increasing use of MALDI has been made in the characterization and detection of disease and in the identification of biomarkers. Some of the glycan biomarkers reported for, for example, cancer, however, appear to be more associated with glycopro-teins involved in inflammation and are, thus, secondary to this disease. Blomme et al. (2009) have noted that "Although individual liver diseases have their own specific markers, the same modifications, hyperfucosylation, increased branching and a bisecting GlcNAc, seem to continuously reappear in all liver diseases." Increases in fucosylated triantennary glycans from AGP is a case in point. Several reviews have appeared and are summarised in Table 22 . Practical details for the characterization by MALDI-TOF and ESI-MS of N-linked glycosylation on recombinant glycoproteins produced in P. pastoris and for detecting potential cancer biomarkers in various cell lines and sera from patients have been published. Bereman, Williams, and Muddiman (2009a) have developed a nano-LC linear trap quadrupole (LTQ) Orbitrap method for analysis of released N-glycans and compared the spectra with those obtained by MALDI-FT-ICR. Whereas the MALDI spectra showed much loss of sialic acids from the sialylated glycans, the Orbitrap spectra showed no decomposition. The method was applied to glycans released from plasma glycoproteins in benign gynecologic tumors and from epithelial ovarian cancer patients. One of the biantennary glycans found to be down-regulated in the cancer patients was a fucosylated biantennary glycan in which the fucose was unusually shown attached to a Gal residue rather than to the core as determined by MS/MS. The compounds were ionized as [MþH] þ species suggesting that this might be an erroneous structure and the result of an internal rearrangement that is known to occur under these conditions. A detailed statistical analysis has been performed on eight data sets of N-glycans released from serum glycoproteins from prostate, breast and ovarian cancer patients (Barkauskas et al., 2009) and measured by MALDI-FT-ICR MS. Significant differences between control and cancer groups were found in all eight datasets. Two structurally related compounds were found to be significantly different between control and cancer groups in all three types of cancer. These compounds had compositions of Hex 3 -HexNAc 4 -Fuc 1 and Hex 5 -HexNAc 4 -Fuc 1 and were probably from IgG rather than being produced by the cancer cells. Narimatsu et al. (2010a) have developed a high-throughput method for detecting cancer biomarkers in early stages of the disease. Briefly, the method consisted of the extraction of tissue mRNAs and measurement of the expression by quantitative realtime polymerase chain reaction (PCR). The results suggested that different glycan structures were synthesized in different cell lines. Secreted proteins from the same cancer cells were collected from serum-free culture and then applied to a lectin microarray to select lectin(s) that showed differential binding to glycoproteins secreted from each cancer cell line. After selection of a specific lectin, isotope-coded glycosylation sitespecific tagging (IGOT) was used to identify core proteins that carry an epitope bound by a specific lectin. Each candidate biomarker was immunoprecipitated from serum using commercially available antibodies and their glycan structures were profiled by lectin microarray, and finally determined by MS n technology with measurements by MALDI-QIT-TOF MS. Other applications are listed in Table 23 . α-Glc(1-7)-α-Hep-(1-5)-α-Kdo4P-(2-6)-β-GlcN4P-(1-6)-α-GlcN1P Arthrobacter globiformis and A. scleromae Diglycosyl glycerol FT-ICR (DHB), NMR Structural determination. Mannose and galactose (Paściak et al., 2010b) Aspergillus fumigatus Glycoinositolphospho-ceramides TOF The mitA gene shown to be required for mannosylation (Kotz et al., 2010) Hymenobacter sp. Carotenoids MALDI Identification of 2'-methyl and 1'-xylosyl derivatives of 2'hydroxyflexixanthin (Klassen et al., 2009) Leishmania major Glycoinositolphospholipids TOF (CHCA), ESI-MS n Demonstration of a UDPglucose independent UDPgalactose salvage pathway (Lamerz et al., 2010) Lipoteichoic acid TOF (DHB) Identification of two enzyme systems involved in biosynthesis Phosphatidyl-myoinositol mannosides (PIMs) Glycopeptidolipids shown to mask PIMS in cell wall (Rhoades et al., 2009) There is currently great interest in the production of therapeutic antibodies and MALDI-TOF MS is frequently used in the analysis of their attached glycans. Reviews have been published on methods for the production and MS analysis of IgG (Huhn et al., 2009) therapeutic antibodies (Beck et al., 2008; del Val, Kontoravdi, & Nagy, 2010; Higgins, 2010; Zhang, Pan, & Chen, 2009i ) and on the humanization of recombinant glycoproteins expressed in insect cells (Tomiya, 2009) . Practical details for characterization of antibody glycans have been published by several investigators Janin-Bussat et al., 2010a ,bJanin-Bussat et al., 2010a . A discussion, with experimental details, of methods based on blot detection with glycan-specific probes, MS of released glycans and LC/MS detection of glycopeptides with the aim of determining whether, how and where plant-derived biopharmaceuticals are glycosylated has also been published . Several new methods have been reported. Thus: A highthroughput method for monitoring IgG glycosylation using a 96well plate format with IgGs purified from 2 mL of human plasma has been developed by Selman et al. (2010) . IgGs were extracted using immobilized protein A, cleaved with trypsin and the resulting glycopeptides were purified by reversed-phase or hydrophilic interaction SPE. Glycopeptides were analyzed by intermediate pressure MALDI-FTICR-MS using both DHB and CHCA, both of which produced signals from sialylated as well as nonsialylated glycopeptides. The method showed an interday variation of below 10% for the six major glycoforms of both IgG1 and IgG2 and was found to be suitable for isotype-specific high throughput IgG glycosylation profiling from human plasma. The method was applied to the IgG glycosylation of 62 human samples. Two lectin-affinity chromatography techniques, Con A and Lens culinaris agglutinin, have been used to enrich, by removal of high-mannose glycans, the nonfucosylated N-glycans from IgG with product detection by MALDI-TOF following PNGase F digestion (Tojo et al., 2009) . Prien et al. (2010) have used a stable isotopically labeled derivative for rapid glycan screening of biotherapeutics. Glycans were labeled with either [ 12 C 6 ]-or 13 C 6 ]-2-AA for both MALDI-TOF or LC-MS analysis. The 2-AA label provided high sensitivity detection in negative ion mode and the mass separation of six units between the isotopically labeled variants eliminated problems arising from isotopic overlap. PEGylation of proteins is frequently used to prolong the serum half-life time of recombinant proteins but their very high MWs put many of them outside the mass range of commercial MALDI-TOF systems using conventional secondary electron multiplier (SEM) detectors. Seyfried et al. (2010) have investigated the use of a high mass (HM) detector combined with a MALDI linear TOF MS system for the detection of PEGylated (glyco)proteins in the range of 60-600 kDa. The system consisted of a Shimadzu AXIMA CFRþ instrument equipped with both a conventional detector and additionally, with an inline moveable HM ion conversion detector (ICD HM1, from CovalX). Spectra were run from sinapinic acid in the linear positive ion mode and were obtained from small (interferon a2a), middle (HSA) and high (coagulation factor VIII and von Willebrand factor (vWF), both heavily glycosylated proteins) molecular mass proteins. The particular challenge was the heterogeneity of the (glyco)proteins in the high MW range in combination with heterogeneity added by the PEGylation, Nevertheless, the performance of MALDI linear TOF MS was found to be superior to that of other methods. Although the SEM was able to obtain information about protein PEGylation in the mass range up to 100 kDa (e.g., PEGylated HSA), the HM system was crucial for detection of ions from the larger compounds, the masses of which sometimes exceeded 0.5 MDa. Detection of these compounds was impossible with the standard SEM. The particular challenge for the analysis was the heterogeneity of the (glyco)proteins in the high MW range in combination with additional PEGylation, which even introduced more heterogeneity and was more challenging for interpretation. Nevertheless, the performance of MALDI linear TOF MS with both detector systems in terms MW and heterogeneity determination depending on the m/z range was superior to the other methods. Isolation and characterization of rhamnolipid-producing bacterial strains from a biodiesel facility (Rooney et al., 2009) Proposed as a member of a new genus and species (Golyshina et al., 2009) Aplysia kurodai (sea hare) Glycosaminoglycans TOF, MS/MS Structural determination (Yoon et al., 2010a) Arthrobacter crystallopoietes N-08 α1,α1-trehalose TOF, ESI, NMR First report that trehalose can be produced from maltose in this bacterium Beverage from fermented plant (Gülcemal et al., 2010) Radix puerariae (Kudzu) TOF Identification from cultivated kudzu root (Nguyen et al., 2009b) Red wine Procyanidins and anthocyanins TOF (indole-3-acrylic acid) Phenolic extracts shown to induce nitric oxide (NO)mediated vasoprotectivity (Auger et al., 2010) Salmo salar ( A novel method for the determination of aminoglycoside antibiotics used surface-assisted laser desorption/ionization mass spectrometry (SALDI MS) with the aid of silver-coated gold nanoparticles (Au@AgNPs) capped by anionic citrate. These nanoparticles were used both as concentrating agents and as matrices in SALDI MS. The LODs at signal-to-noise ratio of 3 were 3, 25, 15, 30, and 38 nM for paromomycin, kanamycin A, neomycin, gentamicin and apramycin respectively. The LODs of the first four of these antibiotics in plasma samples were 9, 130, 81, and 180 nM respectively. Recoveries of the antibiotics from plasma were about 80% (Wang et al., 2009h) . Further examples of the use of MALDI MS in the analysis of therapeutics are given in Table 24 . Applications in this section mainly involve the use of MALDI to investigate products of newly isolated enzymes. These are summarised in Table 25 . Other studies are aimed at elucidating enzyme activity as illustrated by the development of a method to determine the cleavage site in small oligomannoses that has been developed by Hekmat et al. (2010) . Enzymatic cleavages were performed in 18 O-labeled water, conditions that introduced 18 O into the anomeric position of the cleaved glycans. Thus, measurements by MALDI-TOF could determine if a product arose from the non-reducing end of the original oligomannose by its incorporation of the label. The addition of a small amount of various ionic liquids has been found to modify the activity and regioselectivity of different immobilized preparations of Rhizomucor miehei lipase that catalyzes the hydrolysis of hexa-O-acetyl lactal in aqueous media (Filice, Guisan, & Palomo, 2010) . The activity of the enzyme GlcNAc-transferase Vb, which transfers GlcNAc to the 6-position of the 6-antenna in N-glycans has been compared with that of GnT-V. One unusual product found after 8 hr was the addition of GlcNAc to the 6-position of both antennae (Alvarez-Manilla et al., 2010a). Relevant reviews on carbohydrate synthesis are summarised in Table 26 Principal component analysis, shows glycobiological differences between normal and cancer. Hybrid, complex glycans (Goetz et al., 2009b) Breast cancer CD98hc clycoprotein PNGase F, TOF/TOF (DHB), complex glycans (per-Me) Identification of the glycoprotein that binds to GalMBP (fragment of mannose-binding protein) (Powlesland et al., 2009) Breast cancer Serum Trypsin, TOF/TOF (DHB), glycopeptides Expression of Helix pomatia lectin binding glycoproteins in women with breast cancer in relationship to their blood group (Welinder et al., 2009) Breast cancer Cells β-Elimination (NH 4 carbamate), R-TOF/TOF (DHB), glycans, MS/MS Use of new release method using glycoblotting and ammonium carbamate for β-elimination, Core 2 O-glycans Breast cancer Serum Set of glycans identified that can be used as biomarkers (Hammoud et al., 2010) (Continued) Changes in 57 glycans . Three N-glycans sufficient for detection with 90% accuracy Hepatocellular carcinoma Serum glycoproteins TOF/TOF (DHB), glycans (per-Me) Analysis using hierarchical clustering analysis. 7 potential glycan markers identified. Man 5 GlcNAc 2 , bi-, tri-antennary (Tang et al., 2010c) Hepatocellular carcinoma Serum and liver glycoproteins of rats PNGase F, R-TOF (DHB), glycans (per-Me) Increase in core-α-1,6fucosylated glycoproteins. Possible biomarker Hepatocellular carcinoma Suggests shiga toxin as potential therapeutic agent (Distler et al., 2009) Enhanced expression of monosialylated triantennary glycans in cancer (Yoon et al., 2010b) Stomach cancer MKN45 cells, serum Automated β-elim. (Matsuno et al., 2007) , TOF (DHB), glycans (phenylhydrazones) MKN45 cells found to express characteristic trisialopolylactosamine-type glycans. Reduced triantennary complex and increased biantennary glycans at Asn-143 in patients Table 29 and general reactions in Table 30 . Methods to change the glycosylation of a glycoprotein are common for recombinant antibiotic production as outlined above. A general method for producing homogeneous glycoproteins with eukaryotic N-glycosylation has been reported and involves the transfer of the Campylobacter jejuni glycosylation machinery to E. coli and production of glycosylated proteins with the key GlcNAc-Asn linkage. The bacterial glycans were then trimmed and remodeled in vitro by enzymatic transglycosylation to give a eukaryotic-type N-glycosylation (Schwarz et al., 2010) . A method for immobilization of unstable membrane-bound enzymes to a commercially available sepharose support for glycan synthesis has been published Ito et al. (2010c) . It involves modification of the protein C-terminus and a transpeptidase reaction by Staphylococcus aureus sortase A (SrtA) has been developed. Recombinant human b1,4-galactosyltranseferase or recombinant H. pylori a1,3-fucosyltransferases were bound with simple aliphatic amino groups displayed on the surface of the solid support and were shown to have the required glycosyltransferase activity. As with previous reviews in this series, two types of compound appear to be particularly suited to MALDI-TOF analysis; namely glycodendrimers and carbohydrate/protein complexes. Lysosomal proteins in mouse model In contrast to previous assumptions, CHO cells shown to be capable of adding antigenic α-Gal residues to N-glycans (Bosques et al., 2010) α1-Antitrypsin (In human AGE1hn cells) TOF To characterize novel human cell line (Northoff et al., 2010) α1-Proteinase inhibitor (In Aspergillus niger) Trypsin, PNGase F, βelimination, TOF/TOF (DHB), glycans (per-Me), GC/MS Production, purification, and characterization, Glycans = high-mannose (large antennae) (Chill et al., 2009) Anti-CD20 IgG1s, anti-CD20 IgG1 rituximab mutant (S239D/S298A I332E), (Human) Removal of fucosylated antibody ingredients from therapeutics shown to elicit high antibody-dependent cellular cytotoxicity in blood by two mechanisms Anti-EGFRxanti-CD3 bispecific IgG ( Model for comparison of release and glycan analysis methods (Triguero, et al., 2010) Centellosides (In Centella asiatica plant cell cultures) TOF To obtain a more efficient production system. α-Amyrin, converted into centellosides by C. asiatica cells Introduction of bisecting GlcNAc suppresses 1→3-fucosylation and xylose attachment to form paucimannosidic glycans (Karg, et al., 2010) α-L-Iduronidase (Human in seeds of Brassica napus and Nicotiana tabacum) TOF (sinapinic), glycoprotein. GC/MS of monosaccharides Attached carboxy-terminal ER-retention motif, SEKDEL, reduces Xyl and Fuc in N-glycans but has little effect on enzyme activity (Galpin et al., 2010) IgA1 and IgA2 (In murine melanoma cells) PNGase F, L-, R-TOF (DHB, THAP), glycans Structural determination. Hybrid, bi-, tri-, tetra-antennary complex. IgG ( Use of glycosidase inhibitor to produce IgG with Man 9 GlcNAc 2 . Effect on xtal structure and effector functions (Crispin et al., 2009a) IgG (Mouse in tobacco BY2 cells) Hydrazine, TOF (DHB), glycans (2-AP) Use of an ER retention signal (KDEL) increased high-mannose glycans but did not eliminate paucimannosidic glycans IgG (Human 29IJ6 in silkworm larva hemolymph) PNGase F, TOF, glycans (2-AP) Produced 8 mg per kg of larvae with recovery of 83.1%. paucimannosidic glycans IgG (In CHO cells) PNGase F, TOF, glycans Production of non-fucosylated glycans by mutation (Glycans -high-Man, biantennary complex) (von Horsten et al., 2010) IgG (In CHO cells) PNGase F, TOF Chromatographic method to enrich nonfucosylated glycans (Tojo, et al., 2009) IgG ( Effect of culture medium components on product yield. Glycans: G1F, G2F (Paul et al., 2009b) A. Synthesis of Multivalent Carbohydrates, Dendrimers, and Glycoclusters An extensive review of dendrimers (Astruc, 2010 #7260) , with a section on glycodendrimers, illustrates the breadth of this subject. Masses of the larger compounds are frequently in the range 20-50 kDa. The largest compound, based on a polyglycerol scaffold contained an estimated 222 mannose residues but the authors had difficulty obtaining a MALDI spectrum because of the high MW. Syntheses frequently involve Huisgen-type click chemistry because high-yield reactions are essential when so many carbohydrate molecules have to be attached. Table 31 lists papers reporting syntheses of glycodendrimers and similar compounds. Among the largest of these compounds to have been reported during the review period has involved conjugation of LPSderived oligosaccharides to diphtheria toxin CRM 197 protein in an attempt to develop a vaccine against invasive meningococcal disease. Conjugates with a mass of 102 kDa were analyzed by MALDI-TOF from sinapinic acid, the most widely used matrix for compounds of this type. Conjugates can be characterized by MALDI-TOF MS but for large molecules the resolving power of most instruments is insufficient to distinguish each product and only a broad peak is observed. Whereas the center of the peak represents the mean copy number of ligands per protein, information on the dispersity of the sample is usually neglected. Patel et al. (2010) have produced a mathematical approach for calculating dispersity. By simply measuring the width at half maximum of the broad peaks that usually arise from carbohydrate-protein complexes and from the unmodified proteins, they were able to calculate the product distribution variance. Furthermore, since the area between AE2s equates to 95% of the total, m AE 2s represents the range within which 95% of adducts exist, this gives a direct measure of dispersity. As one example of the type of work involved with this type of compound, the glycosylation sites of O-specific polysaccharide of Vibrio cholerae O1, serotype Ogawa linked to BSA via squaric acid chemistry have been determined by MALDI-TOF/ TOF MS (from CHCA). The spectra showed the presence of hapten-BSA neoglycoconjugates with different hapten-BSA ratios (4.3, 6.6, and 13.2). Sites of glycation were determined by comparison of the masses of the peptides resulting from the digestion of the BSA glycoconjugates and BSA itself using tandem MS/MS with a high-collision energy cell. The spectra Improved secretion of molecular chaperone-assisted IgG. No alterations in N-glycans (Dojima, et al., 2010) Interferon-γ, (Human in CHO cells) TOF, glycans (per-Me) Study of intracellular glycosylation activities. Effects of nucleotide sugar precursor feeding (Wong et al., 2010b) Interferon ( Enzyme, which usually fucosylates core of N-glycans, shown to fucosylated chitooligosaccharides (Ihara et al., 2010a) Galactofuranosyltransferase GlfT2 (Recombinant in M. tuberculosis H37Rv) L-, R-TOF (CHCA) Polymer length shown to be controlled by a template-independent polymerase (May et al., 2009) β-D-Galactoside-α1,4-Gal-transferase and β-D-Galactoside-β1,4-Galtransferase from pigeon Two novel enzymes catalyzing the formation of Galα1→4Galβ1→-Galβ1-4GlcNAc GalNAc transferase 2 (human) TOF/TOF (DHB) Adds GalNAc to several unnatural peptides (Yoshimura et al., 2010) GalNAc transferase 10 (human) TOF (DHB) Enzyme found to have a unique GalNAc-O-Ser/Thr-binding site (Perrine et al., 2009) GalNAc transferase 20 (human) TOF Found in testis and brain (Peng et Identification of Gal-transferase adding Gal to core Fuc. (Titz et al., 2009) GnT-I fuzed to maltose binding protein from Nicotiana tabacum TOF/TOF Recombinant expression and characterization (Dohi et al., 2010) GnT-V and GnT-Vb (recombinant human. Incubations in 293T cells) MALDI Enzymes transfer GlcNAc to 6position of 6-antennae in N-glycans. Comparisons of enzymes (Alvarez-Manilla, et al., 2010a) GPI-modifying β1-3-GlcNAc transferase from Trypanosoma brucei Enzyme characterization (Izquierdo et al., 2009) Glycosidases Penicillium canescens TOF (DHB) Isolation and biochemical characterization, (Burtseva et al., 2010) Cellulase Cel45A from Trichoderma reesei TOF/TOF (DHB, THAP), ESI-MS/MS Enzyme shown to catalyze hydrolysis of glucosidic bonds adjacent to mono-substituted anhydroglucose units (Enebro et al., 2009b) Chitinase-A from Cycas revolute (cycad) TOF (DHB) Biochemical characterization, cDNA isolation, and posttranslational modification (Taira et al., 2009b) Chitinase-A and B from Serratia marcescens strain BJL200 expressed in E. coli TOP10 Effects on inhibition efficacy of allosamidin, a general competitive inhibitor of family 18 chitinases. (Zakariassen et al., 2010) Chitinase and N-acetylhexosaminidase from Verticillium fungicola TOF (DHB) effect of pH on the production of enzymes in submerged cultures Cellulomonas fimi TOF (DHB) Use of mutations to affect mannose binding (Hekmat, et al., 2010 ) Exocellulase Cel6B from Thermobifida fusca TOF Proposal of a novel hydrolysis mechanism involving proton-transfer ER Glucosidase II from rat liver TOF Found to be a broad specificity hexosidase (Miyagawa et al., 2010) Family GH38 α-mannosidase from Streptococcus pyogenes TOF, high-Man glycans (per-Me) Characterization. α1→3mannosidase (Suits et al., 2010) β-Galactosidases from Arabidopsis subfamily a1 TOF Comparative characterization (Gantulga et al., 2009) (Continued) β-Galactosidases 4 and 5 from tomato TOF Enzymatic activity and substrate specifcity (Ishimaru et al., 2009) β-Galactosidases (BgaC protein) from Streptococcus pneumoniae Specific for Galβ1-3GlcNAc moiety (Jeong et al., 2009) α-Glucosidase from Aspergillus niger R-TOF (DHB) Monitoring hydrolysis and transglycosylation activity (Shimba et al., 2009) α-Glucosidase from Aspergillus niger TOF (THAP) Activity towards dextrin and starch (Ota et al., 2009) β-Glucuronidase from Aspergillus niger TOF Isolation and substrate specificity for transglycosylation (Kiryu et al., 2009) Shown to be involved in the maintenance of chronic infections (Domenech et al., 2009) Cholinephospho-transferase from Mycoplasma fermentans TOF/TOF (DHB) Molecular cloning and expression of enzyme involved in glycol-glycerophospholipid biosynthesis Hemicellulase from Chrysosporium lucknowense C1 TOF Investigation of the fungus for hemicellulase production (Hinz et al., 2009) showed the presence of three conjugation sites on Lys residues 235, 437, and 455, assumed to be the most accessible. The identification of y-series product ions was found to be useful for sequencing of various peptides and the a-and b-product ions confirmed the sequence of the conjugated peptides . Other examples are listed in Table 32 . MALDI-TOF analysis, combined with IR spectroscopy has demonstrated covalent modifications of chitin with silk-like proteins in the formation of shells in the mollusc Mytilus galloprovincialis (Weiss et al., 2009) . MALDI-TOF MS has also been used as a standard against which to evaluate and optimize a Fluorophore-assisted carbohydrate electrophoresis (FACE) method for analysis of pectic oligosaccharides (Sun et al., 2009a) (Article in Chinese). Koroleva et al. (2010) have used MALDI-TOF MS to identify all the major glucosinolates that accumulate in S-cells of Arabidopsis leaves and flower stalks. Cell sap was diluted in 10 mL methanol and mixed 1:1 with CHCA before being spotted onto the MALDI target plate. The analysis was performed in negative-ion mode using an Ultraflex TOF/TOF instrument. Finally, Mun, Rho, and Kim (2009) have used MALDI to confirm the molecular sizes of commercial cycloamyloses and Patsos et al. (2009) have used it to detect aryl glycans in cells treated with inhibitors of O-glycan processing. MALDI continues to be a major technique for carbohydrate analysis although electrospray is more widely used. A major advantage of MALDI is that is gives a cleaner profile of glycan mixtures because of the absence of multiple charging. New techniques, such as ion mobility have emerged to complement both MALDI and electrospray but there is still much scope for improvement in carbohydrate analysis. Surveys during the review period have highlighted the fact that many laboratories still make mistakes when assigning structures. Much of this can be attributed to assumptions made between a simple mass (Urch et al., 2009) S-Adenosyl-L-methionine dependent methyltransferase from Haloferax volcanii TOF (CHCA) Enzyme shown to methylate N-linked pentasaccharide Tlmk from Strepto-alloteichus hindustanus FT-ICR Functional characterization in tallysomycin biosynthetic pathway (Wang et al., 2009g) Trehalose synthase from Thermotoga maritima DSM3109 TOF Molecular cloning and characterization (Ryu et al., 2010) UDP-arabinopyranose mutase from rice TOF (sinapinic), glycoprotein Arg residue shown to be reversibly glycosylated with single glycosyl residue; residue needed for activity (Konishi et al., 2010b) (Gao et al., 2009b) Aminated xyloglucan (from tamarind) TOF (DHB) Physico-chemical properties of aminated xyloglucan extracted from tamarind seed (Simi & Abraham, 2010c) Bridged C-furanosides TOF Intramolecular nucleophilic attack of BzO group in a triflated cyclooctenol (Jürs & Thiem, 2009 ) 1-Deoxynojirimycins with dansyl capped N-substituents TOF As probes for Morbus Gaucher affected cell lines (Fröhlich et al., 2010) 2,3-Dibromo-3-methyl-1phenylphospholane 1-oxide TOF As novel anticancer agent (Yamada et al., 2010b) Synthesis of polysaccharide intracellular adhesins using an acid reversion reaction of N-acetylglucosamine in HF-pyridine (Leung et al., 2009) Amino-bridged oligosaccharide mimetics R-TOF (DHB), FAB Synthesis using reductive amination. Glycomimetic target structures as potential ligands for the receptor protein NKR P1 of natural killer cells (Neumann & Thiem, 2010) Amylose chains TOF From starch by action of phosphorylase. Preparation of A-type crystals (Montesanti et al., 2010) Bi-fluorescently-labeled maltooligosaccharides TOF For amylase assays (Oka et al., 2010a) Bis-hydrazides TOF For conjugation with proteins etc. (Adak et al., 2010) Blood group tetrasaccharides B (types 1, 3 and 4) TOF 3-Aminopropyl glycosides of tetrasaccharides synthesised using acetylated Galα(1→3)-(Fucα(1→2))Gal trichloroacetimidate as a glycosyl donor (Korchagina et al., 2009) 3,6-Branched arabinogalactan-type tetraand hexa-saccharides TOF For investigation of monoclonal antibodies raised against arabinogalactan proteins from pressed juice of Echinacea purpurea. Carboxymethylated cyclosophoraose TOF As a novel chiral additive for the stereoisomeric separation of flavonoids by CE (Jeon et al., 2010) Chitooligosaccharides TOF By the enzymatic hydrolysis of chitosan ) Chitooligosaccharides TOF By the enzymatic hydrolysis of chitosan (Xu et al., 2010a) Chitooligosaccharides TOF/TOF (DHB) glycans (AMAC) Characterization of family 46 chitosanase from Streptomyces coelicolor A3(2) and use for degradation of chitosans Chitooligosaccharides TOF To study the antibacterial activity against bifidobacteria (Šimůnek et al., 2010) Chitosan and chitooligosaccharides Adsorption properties for uranium (Paper in Chinese) (Jiang et al., 2010b) Chitosans TOF Synthesis from lobster chitin by NaOH deacetylation and enzymatic hydrolysis. To protect crops from main pathogens (Falcón et al., 2010) Chito-tetrasaccharide β-1,4-GlcNAc-β-1,4-GlcN repeat TOF (DHB) By condensation of two disaccharides Deacetylated chitohexaose TOF Hydrolysis of chitosan. Could limit cell proliferation of breast cancer cells (Xiong et al., 2009 ) 2-Deoxy cyclic and linear oligosaccharides TOF (DHB) Synthesis by oligomerization of monomers (Paul et al., 2009a) β-D-Fructopyranosyl-(2→6)-D-glucopyranose TOF Synthesis from D-glucose and D-fructose by thermal treatment (Yamamori et al., 2010) Galactofuranose oligomers TOF/TOF (CHCA) To probe mechanism by which polymer length is controlled in mycobacteria (Splain & Kiessling, 2010) Galactomanno oligosaccharides TOF From hydrolysis of guar gum by βmannosidase (Paper in Chinese) (Zhao, et al., 2009b) Galactooligosaccharides FT-MS, GC/MS Synthesis by acid hydrolysis of the polysaccharides from Nerium indicum Mill α-Glucan pentasaccharide from Aconitum carmichaeli Use of chiral-auxiliary-mediated 1,2-cisglycosylations for the solid-supported synthesis (Boltje et al., 2010) Glucan with α -(1→6) linkages and α -(1→3) and α -(1→4) branch points Produced from glucan of Leuconostoc mesenteroides NRRL B-742 by microwave assisted hydrolysis (Majumder et al., 2009) Glucosylated raffinose TOF (DHB) Use of glucansucrase (alternansucrase) for synthesis Glc-Glc, Gal-Gal, Gal-Glc, Gal-Gal disaccharides TOF (DHB) 16-member library containing all regioisomers. Solid-phase synthesis (Ágoston et al., 2009b) Linear isomaltooligosaccharides (DP2-10) TOF One-step synthesis using synthetic fusion enzyme of dextransucrase and dextranase (Kim et al., 2009e) Macrocyclic oligosaccharides TOF Copper(I)-catalyzed Huisgen's 1,3-dipolar cycloaddition of alkyne and azide provides size-defined macrocyclic carbohydrates (Muthana et al., 2009) Mannose-capped disaccharide with a thiol terminus To provide a tethered sugar for attaching to gold nanoparticles to mimic carbohydrateinvolved cell-surface interactions (Continued) Chemical stereochemically controlled synthesis (Pastore et al., 2010a) Nigerose-containing oligosaccharide TOF/TOF (CHCA) Escherichia coli Non-glycosidically linked pseudodisaccharides TOF (THAP) Thioethers, sulfoxides, sulfones, ethers, selenoethers, and their binding to lectins Oligosaccharides from red seaweed polysaccharides TOF (CHCA), ESI Efficient conversion of galactans into Cglycosyl aldehydes (Ducatti et al., 2009) Oligosaccharide mimics of sialyl Lewis A Trisaccharide. Neu5Ac and Fuc replaced with HSO 3 and D-Ara respectivly (Jakab et al., 2010) Pentasaccharide anticoagulant (Idraparinux) MALDI Compound is fully O-sulfated, O-Me mimic of antithrombin III binding domain of heparin ) N-Quaternary chitosan derivatives TOF Synthesis, characterization and antibacterial activity (Rúnarsson et al., 2010) Sialylated lactosides TOF (DHB), per-Me For coupling to BSA by Huisgen reaction. As glycocongugate antigen (Mosley et al., 2010) Sialyllacto-N-tetraose and sialyllacto-N-neotetraose TOF Use of α2-3and α2-6-sialylated lactosaminide precursors obtained by enzymatic procedures with glycosylations employing triflic acid/N-iodosuccinimide (Schmidt & Thiem, 2010) Sucrose-based guanidinecontaining G7 molecular transporters TOF Show different patterns of intracellular localization depending on the nature of the linker chains as well as the fluorescent dyes (Lee et al., 2009l) Sulfated oligofucosides MALDI Synthesis of sulfated octyl tetra-to octaoligofucosides with different sulfation patterns (Zong et al., 2010a) α To achieve steric instead of electrostatic stabilization. Two-step synthesis (Kloser & Gray, 2010) Cyclic β-glucan R-TOF/TOF (CHCA) Synthesis using laminarinase 16A glycosynthase mutant from the basidiomycete Phanerochaete chrysosporium (Vasur et al., 2010) Dextrin-hydroxyethylmethacrylate and dextrinvinyl acrylate hydrogels TOF/TOF (DHB) For the determination of biocompatibility and biodegradability in mice (Moreira et al., 2010) Epoxy-starch derivatives TOF (DHB) Synthesis by epoxidation of allylated starch (Huijbrechts et al., 2010) Poly-N-acetyllactosamine TOF (DHB, CHCA) Chemo-enzymatic synthesis. Characterization for CGL2-galectin-mediated binding of ECM glycoproteins to biomaterial surfaces (Sauerzapfe et al., 2009) Triblock co-oligomers of tri-O-methylated and unmodified cello-oligosaccharides: TOF (DHB) Synthesis and structure-solubility relationships (Kamitakahara & Nakatsubo, 2010) Xylan-based polysaccharides TOF (DHB) Amino-modified low MW xylan reacted with unmodified xylan and cellodextrins ) Glycosaminoglycans and related compounds N-Acetyl-heparosan oligosaccharides TOF Digestion of N-acetyl-heparosan with heparitinase. for study of enzymology (Sugiura et al., 2010a) 2,3-Desulfated heparin MALDI For control of inflammation by inhibition of E-selectin (Lakshmi et al., 2010) Heparan sulfate oligosaccharides TOF/TOF (DHB), ESI Use of modular building blocks for preparation of a library of 12 oligosaccharides Used to probe the structural features of HS for inhibiting the protease, BACE-1 (Arungundram et al., 2009) Heparan sulfate oligosaccharides TOF Synthesis and inhibition of endothelial cell functions essential for angiogenesis (Cole et al., 2010) Hyaluronan TOF For NMR study of chemical proton exchange over the β(1→3) glycosidic linkage (Nestor et al., 2010) Hyaluronic acid decasaccharide MALDI Chemical synthesis using preactivation-based chemoselective glycosylation strategy Hyaluronic acid subunit and fully protected oligomers MALDI Tetra-, hexa-and octa-saccharides. Multigram synthesis (Virlouvet et al., 2010) Isosteric sulfonate analogues of AT-III binding domain of heparin TOF (THAP) D-GlcA-and L-IdoA-containing disaccharide. related to antithrombin-binding pentasaccharide of heparin. One sulfate ester replaced by Na sulfonato-Me moiety Bisected octasaccharide MALDI Chemical synthesis (Wang et al., 2009e ) Galβ(1→3)[NeuAcα(2→6)] GlcNAcβ(1→2)Man motif TOF Chemical synthesis as molecular probe (Bao et al., 2010) Glucuronyl oligosaccharides TOF, glycans (2-AP) Synthesis of glucuronyl and sulfoglucuronyl oligosaccharides from HNK-1 glycoprotein (Yagi et al., 2008) High-mannose glycans TOF (DHB) Related to HIV gp120. One-pot catalytic glycosidation/Fmoc removal (Pastore et al., 2010b) High-mannose glycans with Glc 3 units TOF (DHB) Study of the conformational properties of the Glc 3 Man unit (Mackeen et al., 2009) High-mannose glycansmethotrexate derivatives TOF Identification of the recognition motif of the glycoprotein-folding sensor enzyme, UDP-Glc: glycoprotein glucosyltransferase Biantennary and high-mannose N-glycans linked to non-reducing terminus of Man 3 GlcNAc 2 core, plus biotin for arrays N-Glycan library TOF (DHB), glycans (2-AP) Enzymatic construction of library for building MS database Phosphorylated highmannose glycans TOF High-mannose glycans from ribonuclease. Incubation with recombinant GlcNAc phosphotransferase. (Bohnsack et al., 2009) (Continued) (Song, et al., 2009f) Sialic acid containing complex-type N-glycan TOF New solid-phase synthesis. Stereoselective βmannosylation, microfluidic α-sialylation and glycosylation of N-PhF 3 acetimidate on JandaJel resin Thiol-terminated nonamannoside TOF For synthesis of microarrays (Dietrich, et al., 2010) Various complex glycans TOF Development of HEK393T expression system for human glycosyltransferases Enzymatic construction of library for building MS database Glycosylated amino acid derived from PSGL-1 TOF Use of trichoroacetimidate donors and thioglycosyl acceptors (Vohra et al., 2009) KL-6 antigen TOF/TOF (DHB) Library of glycans to determine binding to anti-MUC1 antibody Aglycoristocetin derivatives containing hydrophobic side chain-substituted cyclobutenedione. Synthesis and anti-influenza activity Androgenic gland hormone of the woodlouse, (Armadillidium vulgare) TOF With various glycans. Showed that thermodynamically most stable form not most active: Result of disulfide pairing (Katayama et al., 2010b) Antifreeze glycopeptide analogues TOF (DHB, CHCA) Microwave-enhanced synthesis and functional studies (Heggemann et al., 2010) β-hFSH with chitobiose units at the natural linkage sites TOF Use of the Sinaÿ radical glycosidation for simultaneous installation of biantennary chains and glycal chemistry to construct the tetrasaccharide core (Nagorny et al., 2009) β-Sheet-rich protein plus GlcNAc at various positions TOF Effect of GlcNAc position on protein folding kinetics and thermodynamics (Price et al., 2010a) Biantennary N-glycans plus peptide TOF/TOF (DHB) Solid-phase peptide synthesis. Glycans linked -NH-CO-CH 2 -S-Peptide (Murase et al., 2009) Bivalent glycopeptide TOF Mannosides linked with squaric acid (Lindhorst et al., 2010a) CD52 Glycopeptide antigens TOF (DHB, CHCA) Chemo-enzymatic synthesis of glycopeptide with N-and O-linked glycans (Huang et al., 2010c ) C-glycosyl β 2 -and β/β 2peptides MALDI Solution-phase synthesis. 3-8 amino acids (Inaba et al., 2009) C-Linked antifreeze glycoprotein analogues TOF (DHB) Effect of the length of the amide-containing side chain between the carbohydrate moiety and the polypeptide backbone influences ice recrystallization inhibition (Tam et al., 2009) C-Mannosylated peptides TOF (DHB, CHCA) Peptides shown to interact with Hsc70 to modulate its signaling in RAW264.7 cells ) Cyclic antifreeze glycopeptides TOF Exhibited antifreeze activity by forming hexagonal-bipyramidal ice crystals (Hachisu et al., 2009) Cyclic neoglycopeptides MALDI (CHCA) For binding to adjacent protein binding sites in wheat germ agglutinin (Schwefel et al., 2010) Dihydrofolate reductase TOF Glycans = GlcNAc, lactose, maltotriose Analysis as tryptic peptides. For study of effects of glycosylation on stability (Tey et al., 2010) Mass Spectrometry Reviews DOI 10.1002/mas Microwave-assisted synthesis. 5(6)-Carboxyfluorescein shown to be stable Fmoc-threonine bearing a core-2 glycan TOF As building block for PSGL-1 via Fmocassisted solid-phase peptide synthesis (Krishnamurthy et al., 2010 ) O-Fucosylated epidermal growth factor-like repeat 12 of mouse notch-1 receptor Chemical synthesis and studies on folding (Hiruma-Shimizu et al., 2010) Galactosylated naringinase TOF (sinapinic) Modification of glycosylation to effect deglycosylation of rhamnosylated drugs (Garnier et al., 2010) Gal-β-3GalNAc-α-Lys 5 TOF Immunogen design for tumor T antigen immuno-targeting (Sendra et al., 2009) Glycopolypeptide-based cholera toxin inhibitors MALDI Effect of peptide charge and glycan linker length on activity (Maheshwari et al., 2010 ) Glycopeptide carrying tetra-N-Ac-lactosamine containing core 2 decasaccharide TOF (DHB) Solid-phase synthesis (Ueki et al., 2010) Glycosaminoglycan-protein linkage tetraosyl peptide moieties of betaglycan TOF To investigate structures that best serve as a hexosamine acceptor for enzymatic glycosyl transfer (Tamura et al., 2010) Glycosulfopeptides from Nterminus of human endoglycan TOF/TOF (THAP) Containing tyrosine sulfate residues and sialyl Lewis x in core 2 O-glycans and bind to human L-selectin (Leppänen et al., 2010) Glycosylated peptoids TOF By on-resin click (Huisgen reaction) glycoconjugation (Norgren et al., 2009 ) Glycosylated cell-penetrating peptide (R6/W3): Ac-RRWWRRWRR-NH 2 TOF One, two, or three galactose(s), with or without a spacer introduced via a triazole link (Dutot et al., 2010) GRGDY grafted to chitosan TOF (CHCA) Linked with sulfosuccinimidyl-6-[4'-azido-2'nitropheny-lamino]hexanoate as drug carrier N-Glycosylated insulin L-TOF (DHB) Addition of three GlcNAc residues at NH 2 groups on peptide. Use of Endo M to transfer glycan to one of them. (Tomabechi et al., 2010a) N-Glycoproteins carrying intact natural N-glycans TOF/TOF (DHB) Enzymatic synthesis of biantennary glycans (Huang et al., 2009b) Glycosylated analogues of glucagon-like peptide 1 TOF, LC-MS To improve proteolytic stability and blood glucose-lowering activity. Sugars = GlcNAc, LacNAc, sialyl LacNAc synth. by chemoenzymatic approaches (Ueda et al., 2009b) Glycosylated neurotensin analogues TOF Containing O-linked β-melibiose and α-TF antigen or β-lactose units linked by a PEG3 spacer. Exhibit subpicomolar anticonvulsant potency in model of epilepsy (Lee et al., 2009f) Glycosylated tetracontapeptide with acidlabile sialyl-T N antigens MALDI (DHB) 20-Residue glycopeptide-α-thioester and 20residue glycopeptide with Cys at N-terminal. Solid phase synthesis; coupled by Cys NCL Ser . O-Glycopeptide mimetics TOF (DHB, CHCA) Aziridine ring opening as regio-and stereoselective access to O-glycosyl amino acids and their transformation into Oglycopeptide mimetics (Schäfer et al., 2009) Heptasaccharide from Campylobacter jejuni plus AcrA61-210 TOF/TOF (CHCA) Development of NMR method for 3D structural determination of glycoproteins using enzymatically synthesised glycoprotein (Slynko et al., 2009) (Continued) 4-OH-proline oligomers Q-TOF (DHB) Compounds form very stable polyproline II helices (Owens et al., 2010) IgA-hinge peptide TOF To study effect of glycosylation on cis/trans isomerization of prolines (by NMR) Man 5-9 GlcNAc 2 -Asn-N-14 C R-TOF (DHB) For micro-method for determining precise oligosaccharidic specificity of mannosebinding lectins (Debray et al., 2009) Mannosylated lysine derivative TOF Bivalent carbohydrate branching unit. Suitable for solid-phase peptide synthesis Chemical synthesis, for enzymatic study in Dictyostelium (see Table 12 ,O-glycans) (Wang, et al., 2009k) S-Linked glycopeptides MALDI Thioglycosylated building blocks prepared from per-Ac sugars via glycosyl iodides in one-pot fashion and used in sub-monomer solid phase strategy (Comegna & De Riccardis, 2009) Synthesis using using the newly developed N-Troc-protected GM3 and GalN intermediates (Komori et al., 2009) Anchor with N-terminal Cys MALDI General method for producing proteins containing a natural GPI anchor using expressed protein ligation (Schumacher et al., 2010) Anchor with unsaturated lipid chains TOF To investigate mechanism of GPI anchoring (Swarts & Guo, 2010 Bearing pyrazolines, isoxazolines, and dihydropyrimidine-2(1H)-thiones. As antibiotics (El-Sayed et al., 2009) Hydroquinone fructoside L-TOF Synthesis using Leuconostoc mesenteroides levansucrase Hydroquinone glucoside L-TOF (DHB) Synthesis using Leuconostoc mesenteroides levansucrase Lobatoside E-related triterpene glycoside MALDI Gold(I)-catalyzed glycosylation with glycosyl ortho-alkynylbenzoates as donors Man-α-(1→6)-Man-O-Octyl analogues MALDI Synthesis and evaluation as potential substrates and inhibitors of a PPM-dependent α-(1→6)-mannosyltransferase involved in LAM/LM biosynthesis (Tam & Lowary, 2010) Mannosylated pyreneperfluoroalkyl lipid MALDI To study multivalent binding on the lateral phase separation of adhesive lipids (Liem et al., 2010) Mannosyl glycolipids with perfluoroalkyl membrane anchors MALDI To assess the cluster glycoside effect during the binding of concanavalin A to mannosylated artificial lipid rafts (Noble et al., 2009) Mesogenic azobenzenes tethered to sugar alcohols (Gretskaya & Mikhalyov, 2007) Cholesterol plus linear glucose TOF (DHB) Synthesis and gelling properties 1,2-Dipalmitoyl-3-(Npalmitoyl-6'-amino-6'-deoxyα-D-glucosyl)-sn-glycerol TOF Glycoglycerolipid of a marine alga with a high inhibitor activity against human Myt1kinase. Synthesis starting from α-Me-Glcp Enzymatic synthesis from 1,2dipalmitoylglycerol in one pot reaction Hydroquinone galactosides L-TOF (DHB) A potential skin whitening agent. Synthesis by reaction of lactase with lactose as donor. Fluorescently labelled synthetic ionophore synthesises in 12 steps with 26% yield (Coppola et al., 2010) Phosphatidyl-myo-inositol mannosides TOF To study mannosyltransferase Corynebacterium glutamicum PimB' (Batt et al., 2010) Puerarin-cycloamylose inclusion complex L-TOF (DHB) Enzymatic synthesis using 4-αglucanotransferase and maltogenic amylase (Choi et al., 2010) Sialic Cinnamoyl-α-cyclodextrin TOF Initiates polymerization of δ-valerolactone (Osaki et al., 2009) Cinnamoyl α-cyclodextrin TOF CDs self-organize to give different supramolecular complexes in aqueous solutions (Tomimasu et al., 2009) (Continued) Synthesized via hydrosilylation reaction under thermal or UV-activated polymerization (Tian et al., 2009) CD-based telluronic acid plus Mn(III)meso-tetra[1-(1adamantyl methyl ketone)-4pyridyl] porphyrin MALDI As artificial enzyme with superoxide dismutase and glutathione peroxidase activities (Yu et al., 2010d) Cyclodextrin dimers TOF One homo-dimer of β-CD and two heterodimers of α-CD and β-CD as hydrolases (Ikeda et al., 2010) Cyclodextrin dimers and trimers TOF/TOF, ESI Bridged CDs with links at different positions formed supramolecular adducts with shapespecific ligands (Aime et al., 2009) β-Cyclodextrin cyclic-nitrone conjugate MALDI With superoxide radical anion and dodecyl chain for membrane insertion α-, βand γ-Cyclodextrinesters (acrylate, pent-4-enoate and undec-10-enoate) Synthesis with nitrophenol esters (Nielsen et al., 2010a) β-CD plus hydroquinone-αglycoside TOF Synthesis and doxorubicin-inclusion abilities (Oda et al., 2009) β-CD monoalkyn TOF (DHB) For conjugation to long-chain thiols for selfassembled monolayer prep. (Dubacheva et al., 2010) Cyclodextrin-polyester polymers TOF Ring-opening polymerization -heating cyclic esters and CDs (Harada, 2009) γ-CDs possessing an azido group and a triisopropylbenzenesulfonyl group As useful synthetic and authentic intermediates for unsymmetrically functionalized derivatives (Himeno et al., 2009) β-CD-ended linear poly(Nisopropylacrylamide) (β-CD-PNIPAM) TOF (THAP, DCTB) Self-assembly of PNIPAM-based amphiphiles formed by inclusion complexation (Zou et al., 2009) β-CD with octadecyl-linked perylene bisimide MALDI Self-assembled amphiphilic perylene-CD conjugate for vapor sensing of organic amines (Jiang et al., 2010a) [6-Deoxy-6-(1-H-1,2,3triazol-4-yl)(Me)6-(4-OMebi-Ph-4'-yloxy) hexanoyl]-βcyclodextrin TOF (DHB) Synthesis, liquid-crystalline properties, and supramolecular organization for controlled release application. (Adrian et al., 2009) Inclusion complexes of β-CD with bipyridine guests TOF With 4,4'-vinyl-enedipyridine, 2,2'vinylenedipyridine, 1-(2-pyridyl)-2-(4pyridyl)ethylene, 4,4'-ethylene-dipyridine, 4,4'-dithiodipyridine, and 2,2'dithiodipyridine Insulated molecular wire with highly conductive πconjugated polymer core TOF Rod-like -Ph-C=C-Ph-core coated with α-CDs (Terao et al., 2009b) Ionic-liquid-functionalized βcyclodextrin TOF As bonded chiral stationary phases for HPLC (Zhou et al., 2010b ) Linear α-cyclodextrin oligomers TOF Controlled synthesis using copper-catalyzed Huisgen 1,3-dipolar cycloaddition (Rawal et al., 2010) Oligothiophene derivatives bearing β-cyclodextrin TOF (DHB) 2T-β-CD 2 and 3T-β-CD 2 , with bithiophene and terthiophene with β-CD at each end form supramolecular assemblies in aqueous solns. (Sakamoto et al., 2009b) Perylene bisimide-bridged bis-(permethyl-β-CDs) FT-ICR As solid-state fluorescence sensor for vapor detection Perylene-bridged bis(βcyclodextrin) FT-ICR (Kitano et al., 2009) Glycodynamers MALDI Dynamic polymers bearing oligosaccharide residues such as Glc 6 obtained from α-CD (Ruff et al., 2010) Membranes (cross-linked) TOF (DHB) Based on acrylated cyclodextrins and polyethylene glycol dimethacrylates (Rölling et al., 2010) Multivalent Galactotrehaloses TOF α,β-GT isomers converted into vinyl monomers then radical copoly-merization with 4-acrylamidophemyl-β-Glc or β-GlcNAc) with acrylamide (Miyachi et al., 2009b) Pentafluorostyrene copolymers with glucose TOF Synthesis by nitroxide-mediated polymerization and "click" chemistry (Becer et al., 2009) Polydioxanone with a protected monosaccharide end-group L-TOF (dithranol) Ring opening polymerization of p-dioxanone using a protected monosaccharide (1,2;3,4-di-O-isopropylidene-α-D-galactopyranose) /Al(OiPr) 3 initiator system (Sugih et al., 2009) Polyfluorene derivative with 20 mol% 2,1,3benzothiadiazole plus α-Man On an amine-reactive hydrogel-coated microarray glass surface. To detect autoantibodies in breast cancer Correction: (Blixt, et al., 2011) Nanofibres TOF Nanofibers formed through π· · ·π stacking of the complexes of glucosyl-C2-salicyl-imine and phenylalanine (Continued) For study of carbohydrate-protein interaction Oligosaccharides from plants TOF (DHB) For construction of microarray to screen for plant transglycosidases activity (Kosík, et al., 2010 ) 6-sulfo-N-acetyl-Dglucosamine containing, on gold TOF To study mechanism of amyloidosis of amyloid β peptides (Fukuda et al., 2010b) Miscellaneous β-D-Allopyranoside-grafted Ru(II) complex TOF Synthesis and both acid-base and DNAbinding properties Azo-sugar nucleotides plus many alkynes TOF (DHB) Click chemistry (Huisgen reaction) to produce inhibitors of glycosyltransferases 1-Deoxy-1-nitropiperidinoses MALDI Synthesis from a protected galactooctopyranose (Collin & Vasella, 2010 ) 4,4-Di-F-5,7-di-Me-4-bora-3a,4a-diaza-sindacene-3propionic acid derivative of Man 5 GlcNAc 2 . (BODIPY) For development of fluorescence assay (Haga et al., 2009) Carbohydrate-functionalized salphen-metal complexes TOF Complexes with peripheral glucose and galactose substituents. Self-assembled supramolecular structures were produced (Hui et al., 2009) Dioxolane-type (9'anthracenyl)methylene acetals TOF Synthesis, regioselective hydrogenolysis, partial hydrogenation, conformational study (Jakab et al., 2009) (Kim et al., 2009f) Amino-sugars TOF (CHCA), ESI Deprotection method for the 2,2,2trichloroethoxycarbonyl (Troc) group using tetrabutylammonium fluoride 1,6-Anhydrosaccharides TOF Use of 7 mol % of AuBr3 to catalyse glycosylations of 6-OH-propargyl/Me mono-, diand tri-saccharides (Thadke & Hotha, 2010) Colored carbohydrates TOF Addition of a Fmoc analogue protecting group based on guaiazulene (Aumüller & Lindhorst, 2009) (Bindschädler et al., 2009) Glycoamino acid building blocks TOF Use of Staudinger ligation Glycopeptides TOF Use of pyruvoyl as a novel protecting group for solid-phase synthesis (Katayam et al., 2009 ) GPI-Anchored proteins TOF Sortase A-catalyzed transpeptidation of GPI derivatives for chemoenzymatic synthesis (Wu et al., 2010f) Heptasaccharide asparagine building block TOF (DHB) Use of one-pot catalytic glycosidation/Fmoc removal (Mezzato & Unverzagt, 2010) Homolinear α(1→6)-linked octamannosyl thioglycosides MALDI Imidazolium cation-tagged mannosyl fluoride and thiomannoside using block couplings. Efficient alternate approach for oligosaccharide synthesis (Yerneni et al., 2009) Homooligosaccharides TOF (DHB) (per-Ac, per-Me) Base-promoted glycosylation of unprotected glycosyl fluorides (Steinmann et al., 2010) Lipophilic thioglycosides TOF Use of heavy lipophilic tag to assist biphasic liquid-liquid separation (Encinas & Chiara, 2009) Macrocyclic neoglycoconjugates TOF (CHCA) Macrocyclization of linear D-galacto-2heptulopyranose-containing oligoketosides by intramolecular glycosidation and ring-closing metathesis Mucin-type Oglycopeptides and oligosaccharides TOF (DHB, THAP, CHCA) Using transglycosylation and reverse-hydrolysis activities of Bifidobacterium endo-α-Nacetylgalactosaminidase Neu5Gc-containing glycans TOF Sialylation with N-glycolylneuraminyl phosphite (Hanashima et al., 2009 ) N-linked glycopeptides TOF Condensation of glycosylamines with Asn. Demonstrated with Man 8 GlcNAc 2 . (Chen & Tolbert, 2010) measurement and structure, particularly when structures are selected directly from databases. At present, no one mass spectrometric technique can identify all structural features of carbohydrates. Sialylated and sulfated glycans still remain a problem although sialylated glycans can be handled after suitable derivatization, particularly by permethylation or simply by methyl ester formation. Permethylation appears to be becoming more popular, particularly for quantification and MALDI. The next few years are expected to bring many developments, particularly in instrumentation and ionization techniques, that will possibly address some of the problems with carbohydrate analysis highlighted above. 2-AB 2-aminobenzamide 2-AA 3-aminobenzoic acid 9-AA 9-aminoacridine ABOE aminobenzoic acid octyl ester AEAB 2-amino-N-(2-aminoethyl)-benzamide AGE advanced glycation end products AGP a1-acid glycoprotein AlPcs aluminum-phthalocyanines a-TF a-Thomsen-Freidenreich antigen All allose AMAC aminoacridone AMT 5-amino-2-mercapto-1,3,4-thiadiazole ANN artificial neural networks ANP 2-amino-5-nitro-4-picoline ANSA 5-amino-2-naphthalenesulfonic acid 2-AP 2-aminopyridine APB aminophenylboronic APCI atmospheric pressure chemical ionization Api apiose APTS 8-aminopyrene-1,3,6-trisulphonic acid AQ aminoquinoline (3-or 5-) Use of genetic engineering to produce glycoproteins in E. coli which are then enzymatically remodelled (see text) (Schwarz, et al., 2010) O-Sulfated trisaccharyl glycopeptide TOF (DHB) Solid-phase synthesis. Use of new benzyl protection method. (Kawahira et al., 2009) Oligosaccharides TOF (DHB) Rate acceleration on stereoselectivity and velocity of O-glycosylation reactions (Ishiwata et al., 2010) Pseudooligosaccharides MALDI Use of cross-metathesis reaction between sugarolefins, followed by intramolecular cyclization (Ronchi et al., 2009 ) S-Linked glycoconjugates TOF, ESI By-thiyl glycosylation of olefinic proteins (Floyd et al., 2009) Sialylated glycans TOF Use of Koenigs-Knorr reaction (with Ag 2 CO 3 ) (Pazynina et al., 2010) Thioglycosides R-TOF/TOF Solvent-free synthesis by use of ball-milling (Patil & Kartha, 2009 ) Trisaccharide libraries TOF Use of linker-tagged building blocks imobilized on a soluble polymeric support (Elsayed, 2009) Acceptor specificity in transglycosylation reactions using Endo-M L-TOF (DHB), ESI (Tomabechi et al., 2010b ) Acetolysis of 6-deoxyhexosemethyl glycosides. Role of sugar configuration TOF (DHB) (Cirillo et al., 2009 ) Acid-catalyzed hydrolysis of β-1,4-glucan, including cellobiose and crystalline cellulose with SO 3 H-bearing amorphous carbon TOF (Suganuma et al., 2010) Allyloxycarbonyl group removal provides a practical orthogonal protective strategy for carbohydrates TOF (Zong et al., 2009 ) Amadori ketoses synthesis in microwave field via Mo VI -catalyzed stereospecific isomerization of 2-C-branched sugars bearing azido function TOF (Hricovíniová, 2010 (Camponovo et al., 2009) Benzhydrylamine-lysine capped with 2 to 64 mono-, di-and tri-α-D-Manp residues TOF Reactive N-OH-succinimide esters to ensure complete reaction of dendrimer amines (G3 mass = 13,841 Da) (Greatrex et al., 2009) 4,7-Bis(9,9-bis(2-(2-(2azidoethoxy)-ethoxy)ethyl)fluorenyl)benzo-thiadiazole with 4 mannose residues TOF (DHB) As an intelligent energy transfer pair for label-free visual detection of concanavalin A Boltorn h30 (commercial polymer) with 27 Gal-Cer groups TOF Binds HIV-1 gp120 Optically pure fullerodendron formed by diastereoselective Diels-Alder reaction Calixarene-based glycocluster oligonucleotide with 4 galactose residues TOF Click chemistry. Triazole-tethered glycoclusters with 3 arrangements. Affinities towards PA-IL and RCA 120 with DNA-based glycoarray. (Moni et al., 2009) Carbosilane with 3, 4 or 6 sialyl-α-(2→3)-lactose residues TOF For anti-influenza properties Carbosilane with 3, 4, 6 or 12 Neu5Ac residues TOF, FAB, ESI Influenza neuraminidase inhibitors. Dendrimers uniformly functionalized with thioglycoside-type sialic acid moieties resistant to neuraminidases (Sakamoto et al., 2009a) Carbosilane with 8 or 16 mannose residues TOF (dithranol) Hydrosilylation of allyl tetra-Ac-Man with carbosilane dendrimers containing monohydrosilane end groups and the subsequent deacetylation (Ortega et al., 2010) Carbosilane with 4 sialyl-Nacetyllactosamine groups TOF Combined chemical and enzymatic synthesis (Matsuoka et al., 2010) Cyclic α-(1→6)-octaglucoside with from 2-7 mannose residues TOF Oxidation of vic-diols, reductive amination with 2-aminoethylmannoside β-Cyclodextrin with 27 mannose or glucose residues TOF (DHB) Heteroglycoclusters. 7 Antennae each with two mannose and one glucose. For lectin binding (Gómez-García et al., 2010) β-Cyclodextrin with dextran TOF (CHCA) Synthesis by click chemistry using Huisgen reaction (Nielsen et al., 2010b) Cyclopeptide with 4 mannose residues TOF Synthesis by click chemistry and molecular recognition study by surface plasmon resonance (Chen et al., 2009c) Dihydroxy-benzamide based with 2-8 mannose, galactose, lactose, glucose or GlcNAc groups TOF (CHCA) Octa-dendrimers. For screening of lectins for multivalency effects. Click chemistry (Pera et al., 2010b) Diphenyldisulfide with 4 glucose residues and others without sugars TOF, GC/MS Use as catalyst to convert allyl alcohols into carbonyl compounds (Tsuboi et al., 2009) Cysteine with 3 or 4 (from dicysteine) mannose residues TOF As inhibitors of type 1 fimbriae mediated bacterial adhesion (Schierholt et al., 2010) Ethylene glycols with 2, 3 or 4 galactose,lactose, maltose or LacNAc groups Ferrocene with one or two glucose, mannose or lactose residues TOF/TOF As electrochemical probes for molecular recognition studies (Casas-Solvas et al., 2009b) Polyglycerol substituted phenylboronic acid TOF Dendrimer formed adducts with, Fru, D-(+)-Gal, D-(+)-Glc, D-(+)-Man, and Me-α-D-Man, by removal of four H 2 O (Hashidzume & Zimmerman, 2009) Fullerene. Sugar balls with 12 iminoglucose residues TOF Synthesis by click chemistry. Glucosidase inhibition shown with resulting iminosugar balls (Compain et al., 2010) Fullerene. Sugar balls with 12 glucose or galactose residues TOF, ESI, FAB 12 residues on 6 arms, Synthesis by click chemistry Gd-diethylenetri-aminepentaacetic acid with 2 glucose residues TOF Synthesis, in vitro and in vivo studies of Gd-DTPA-XDA-D1-Glc(OH) complex as a new potential MRI contrast agent (Ozaki et al., 2010a) Hydroxy benzenes and naphthalenes with 2, 4 or 8 mannose residues TOF Synthesis by click chemistry (Rajakumar et al., 2009 ) L-Lysine plus Gd chelates and 4 galactose residues TOF/TOF As liver imaging probes (Luo et al., 2009) Tetramer of glycodecapeptide from MUC1 with sialyl T N antigen as vaccine candidate (Keil et al., 2009) Peptide/tris-OH-methylmethylamine. 9 galactose or mannose residues attached to β-CD containing doxorubicin TOF For uptake studies in the human hepatocellular carcinoma cell line HepG2 (Bernardes et al., 2010) Peptide/tris-OH-Me-methylamine/ [Ru(bipy) 3 ] 2+ with 18 galactose or mannose residues MALDI To study lectin interactions by monitoring change in fluorescence quantum yield of Ru(II). (Kikkeri et al., 2010a) Phloroglucinol, triazyne, tetrachlorosilane, pentaerythritol, myo-inositol with 2, 3, 4 or 6 Clinked sialic acids TOF Synthesis by Huisgen cycloaddition of azide and alkyne (click chemistry). To explore sialic acid binding to cell surfaces (Papin et al., 2009) Phthalocyanine with 4 α-Galp residues TOF Potential application as photosensitizers in photodynamic therapy (Soares et al., 2009) Pentaerythritol, methyl α-Dglucopyranoside, D-glucose, and Dmannitol with 4, 5 or 6 mannose TOF (DHB) Synthesis by click chemistry, Measurement of binding affinities (Sattin et al., 2010) Poly-glycerol with 8 mannose residues TOF MALDI poor because of high MW (up to 493 kDa) (Kizhakkedathu et al., 2010) Poly-lysine with 16 biantennary Nglycans TOF Synthesis by click chemistry. Effect of sialic acid linkage on in vivo dynamics. Mass around 40 kDa (Tanaka et al., 2010b) Poly-lysine on tris-(2-ethylamino)amine with 3 aulfated cellobiose groups TOF (sDHB) Synthesized by sulfation of polylysinedendritic cellobiose (prepared from cellobiose and polylysine dendrimer generation 3) (Han et al., 2010b) Porphyrin with 8 lactose glycans TOF Synthesis by Huisgen click cycloaddition of azide and alkyne (Okada et al., 2009b) Porphyrin with galactose or lactose MALDI Synthesis by click chemistry. Activity on carcinogenic HEp2 cells Activators of natural killer lymphocytes (Renaudet et al., 2010) Ruthenium(II) bipyridine with 6 or 18 mannose or galactose residues TOF As probes to study lectin-carbohydrate interactions and to measure mono and oligo-saccharide concentrations electrochemically (Kikkeri et al., 2010b) Ruthenium porphyrins with 4 glucose residues TOF Water-soluble catalyst for carbenoid transfer reactions Tetrabenzo-porphyrins with 4 glucose residues TOF To improve targeting of cancer cells (Ménard et al., 2009) Tetraphenyl-ethylene with 4 lactose or sialyl-lactose residues TOF/TOF Synthesis by click chemistry. As fluorescent probes for detection of influenza virus Tetraphenyl-ethylene with 4 or 8 mannose residues TOF (DHB) For fluorescence turn-on sensing of lectins based on aggregation-induced emission (Sanji et al., 2010) Dihydroxybenzamide base with 4 galabiose residues TOF (CHCA) For isolation of pathogenic Streptococcus suis bacteria (Pera et al., 2010a) 1,3,5-Tris-(2-propynyloxy)-benzene with 3 β-cyclodextrin rings TOF (DHB) Synthesis by microwave-assisted click chemistry as fluorescent tripod detection system for pesticides (Mallard-Favier et al., 2009) Trimesic acid and others with 2, 3 or 4 phosphocholine residue related to glycol-sphingolipid from earthworm Pheretima hilgendorfi TOF For enhanced immune responses when compared to their monovalent counterparts Various amino-alkyl with 2, 3 or 4 lactose residues TOF (CHCA) Synthesis from carbamate-linked lactose ) Zinc(II) phthalocyanines with 8 Glc, Gal or cellobiose residues TOF (DHB) Chemical synthesis as photosensitizer in photodynamic therapy (Iqbal et al., 2009a ) Zinc(II) phthalocyanines with 8 Glc, Gal, cellobiose or maltose residues TOF (DHB, CHCA) Eight residues. For photodynamic therapy (Iqbal et al., 2010 ) Zinc(II) naphthalocyanines with 4 glucose residues TOF As photosensitizer in photodynamic therapy (Iqbal et al., 2009b ) Zinc(II) naphthalocyanine with 8 glucose or galactose residues TOF, ESI Ex post glycoconjugation (Berthold et al., 2010) Free amino group on LPS used for site-specific conjugation. Towards bivalent immunogens (Grandjean et al., 2009) 4-Amino-4-deoxy-Larabinose (Ara4N) maleimideactivated BSA Potent immunogen (Müller et al., 2010a) N-Acyl-modified sialylated glycans HSA TOF Squaric acid chemistry. As inhibitors of adenoviruses causing epidemic keratoconjunctivitis Biantennary Gal, 2-keto-Gal + fluorescent probe Single-chain antibody TOF Method for drug targeting using antibodies Galactose BSA TOF New photoinduced thiol-ene coupling Galactose HSA TOF Two step synthesis of Gal 28 as optical imaging agent for peritoneal carcinomatosis (Regino et al., 2010) Ginsenoside Rg3 BSA TOF (sinapinic) Generation and characterization of monoclonal antibody to ginsenoside Rg3 (Joo et al., 2009) α-Glc, α-Man, β-Gal, α- To study role on antigen in immune system interaction (Tefsen et al., 2009) Heparin tetra-saccharide Complement factor H TOF (sinapinic, CHCA) Study of interaction between protein and tetrasaccharide by cross-linking (Blaum et al., 2010) (Hirata et al., 2010) α-Mannosides (bi-and penta-) BSA TOF Immunogenicity and induction of candidacidal activity Methyl glyoxal (advanced glycation end product, Polysialic acid Insulin MALDI Shown to increase insulin lifetime in vivo (Bezuglov et al., 2009) Serogroup 6 pneumococcal oligosaccharides BSA SELDI-TOF, FAB Synthetic carbohydrate conjugates express epitopes found in native capsular polysaccharides (Parameswar et al., 2009) Tn antigen HSA MALDI To study effects of hapten density on induced antibody repertoire Tumor-associated MUC1 glycopeptides To study multivalent binding to human α-defensin (HD5) (Lehrer et al., 2009) FAIMS field asymmetric waveform ion mobility spectrometry Fc fragment (crystallisable) region of IgG Fmoc 9-fluorenylmethoxycarbonyl Fru fructose F-SPE fluorous solid phase extraction FT Fourier transform Fuc fucose G0 (G1, G2) biantennary glycans with 0, (1 or 2) galactose residues G3CA coumaric 1,1,3,3,-tetra-methylguanidine GABA gamma-aminobutyric acid GAGS glycosaminoglycans Gal galactose GalA galacturonic acid GalN galactosamine GalNAc N-acetylgalactosamine G3CA coumaric 1,1,3,3,-tetra-methylguanidine (liquid matrix) GaPcs gallium-phthalocyanines GC/MS gas chromatography/mass spectrometry Production of chitooligosaccharides and their potential applications in medicine Synthesis of biotinylated sialoside to probe CD22-ligand interactions Nanofibers formed through p…p stacking of the complexes of glucosyl-C2-salicyl-imine and phenylalanine: Characterization by microscopy, modeling by molecular mechanics, and interaction by a-helical and b-sheet proteins Bishydrazide glycoconjugates for lectin recognition and capture of bacterial pathogens The applicability of enzymes in cellulose ether analysis Electron impact ion fragmentation pathways of peracetylated C-glycoside ketones derived from cyclic 1,3-diketones Inclusion complexes of gcyclodextrin and carboxyl-modified g-cyclodextrin with C60: Synthesis, characterization and controlled release application via microgels Biochemical characterization of two xylanases from yeast Pseudozyma hubeiensis producing only xylooligosaccharides Synthesis of 4-O-glycosylated 1,5-anhydro-D-fructose and of 1,5-anhydro-D-tagatose from a common intermediate 2,3-O-isopropylidene-D-fructose Solid-phase random glycosylation Comparative solution and solid-phase glycosylations toward a disaccharide library Mutational and functional analysis of Large in a novel CHO glycosylation mutant Separation of 2-aminobenzamide labeled glycans using hydrophilic interaction chromatography columns packed with 1.7 mm sorbent Depolymerization of sodium alginate under hydrothermal conditions Cell wall b-(1,6)-glucan of Saccharomyces cerevisiae. Structural characterization and in situ synthesis New cyclodextrin dimers and trimers capable of forming supramolecular adducts with shape-specific ligands Development of a mouse monoclonal antibody against the chondroitin sulfate-protein linkage region derived from shark cartilage Reflection colour changes in cholesteric liquid crystals after the addition and photochemical isomerization of mesogenic azobenzenes tethered to sugar alcohols Preparative enzymatic synthesis of polyprenyl-pyrophosphoryl-N-acetylglucosamine, an essential lipid intermediate for the biosynthesis of various bacterial cell envelope polymers Plant cell wall proteomics: Mass spectrometry data, a trove for research on protein structure/function relationships Introducing capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) for the characterization of konjac glucomannan oligosaccharides and their in vitro fermentation behavior Introducing capillary electrophoresis with laser-induced fluorescence (CE-LIF) as a potential analysis and quantification tool for galactooligosaccharides extracted from complex food matrices NMR spectral mapping of Lipid A molecular patterns affected by interaction with the innate immune receptor CD14 Bioconjugation of D-glucuronic acid sodium salt to well-defined primary amine-containing homopolymers and block copolymers Glycomic analysis of sialic acid linkages in glycans derived from blood serum glycoproteins Chip-based reversed-phase liquid chromatography-mass spectrometry of permethylated N-linked glycans: A potential methodology for cancerbiomarker discovery Plasticity of xyloglucan composition in bean (Phaseolus vulgaris)-cultured cells during habituation and dehabituation to lethal concentrations of dichlobenil Comparison of the substrate specificities and catalytic properties of the sister N-acetylglucosaminyltransferases, GnT-Vand GnT-Vb (IX) Glycoproteomic analysis of embryonic stem cells: Identification of potential glycobiomarkers using lectin affinity chromatography of glycopeptides Negative-ion MALDI-QIT-TOFMS n for structural determination of fucosylated and sialylated oligosaccharides labeled with a pyrene derivative Structural determination by negative-ion MALDI-QIT-TOFMS n after pyrene derivatization of variously fucosylated oligosaccharides with branched decaose cores from human milk Derivatization with 1-pyrenyldiazomethane enhances ionization of glycopeptides but not peptides in matrix-assisted laser desorption/ionization mass spectrometry Large-scale glycomics for discovering cancer-associated N-glycans by integrating glycoblotting and mass spectrometry A concise review of mass spectrometry imaging A glycomics approach to the discovery of potential cancer biomarkers Determination of glycosylation sites and site-specific heterogeneity in glycoproteins Glycomics and disease markers Structural analysis of a fucoidan from the brown alga Fucus evanescens by MALDI-TOF and tandem ESI mass spectrometry Structural analysis of a highly sulfated fucan from the brown alga Laminaria cichorioides by tandem MALDI and ESI mass spectrometry N-Glycan targeted gene delivery to the dendritic cell SIGN receptor Oxazole-modified glycopeptides that target arthritis-associated class II MHC Aq and DR4 proteins Glycocluster design for improved avidity and selectivity in blocking human lectin/plant toxin binding to glycoproteins and cells Carbamate-linked lactose: Design of clusters and evidence for selectivity to block binding of human lectins to (neo)glycoproteins with increasing degree of branching and to tumor cells MALDI-TOF/MS analysis of archaebacterial lipids in lyophilized membranes dry-mixed with 9-aminoacridine Structural characterization of cell wall polysaccharides from two plant species endemic to central Africa, Fleurya aestuans and Phragmenthera capitata Disease-associated glycosylated molecular variants of human C-reactive protein activate complement-mediated hemolysis of erythrocytes in tuberculosis and Indian visceral leishmaniasis The N domain of human angiotensin-I-converting enzyme: The role of N-glycosylation and the crystal structure in complex with an N domain-specific phosphinic inhibitor, RXP407 Loss of effector function of human cytolytic T lymphocytes is accompanied by major alterations in N-and Oglycosylation Glycome informatics: Methods and applications 5a-Carba-glycopyranoside primers: Potential building blocks for biocombinatorial synthesis of glycosphingolipid analogues Identification, characterization and immunogenicity of an O-antigen capsular polysaccharide of Francisella tularensis Characterization of oligomeric xylan structures from corn fiber resistant to pretreatment and simultaneous saccharification and fermentation Dense-shell glycodendrimers: UV/Vis and electron, paramagnetic resonance study of metal ion complexation Bioinformatics in glycomics: Glycan characterization with mass spectrometric data using SimGlycan TM Functional identification of the Proteus mirabilis core lipopolysaccharide biosynthesis genes Three enzymatic steps required for the galactosamine incorporation into core lipopolysaccharide Redox proteomics of fat globules unveils broad protein lactosylation and compositional changes in milk samples subjected to various technological procedures Lactosomes: Structural and compositional classification of unique nanometer-sized protein lipid particles of human milk The Pleurotus ostreatus hydrophobin Vmh2 and its interaction with glucans Comparison of the water-soluble carbohydrate composition and fructan structures of Agave tequilana plants of different ages Synthesis, conformation, and biological characterization of a sugar derivative of morphine that is a potent, long-lasting, and nontolerant antinociceptive Modular synthesis of heparan sulfate oligosaccharides for structure-activity relationship studies Direct profiling of saccharides, organic acids and anthocyanins in fruits using electrospray droplet impact/secondary ion mass spectrometry Syntheses of mucin-type O-glycopeptides and oligosaccharides using transglycosylation and reverse-hydrolysis activities of Bifidobacterium endo-a-Nacetylgalactosaminidase Recovery of water-soluble compounds from Ganoderma lucidum by hydrothermal treatment Binding and cellular activation studies reveal that toll-like receptor 2 can differentially recognize peptidoglycan from gram-positive and gram-negative bacteria Grafting of aminated oligogalacturonans onto Douglas fir barks. A new ANALYSIS OF CARBOHYDRATES AND GLYCOCONJUGATES & route for the enhancement of their lead (II) binding capacities Defining criteria for oligomannose immunogens for HIV using icosahedral virus capsid scaffolds Dendrimers designed for functions: From physical, photophysical, and supramolecular properties to applications in sensing, catalysis, molecular electronics, photonics, and nanomedicine The red wine extract-induced activation of endothelial nitric oxide synthase is mediated by a great variety of polyphenolic compounds Coloring carbohydrates: Investigation of azulene derivatives as blue protecting groups Differences in the sialylation patterns of membrane stress proteins in chemical carcinogen-induced tumors developed in BALB/c and IL-1a deficient mice Mass spectrometry of N-linked glycans Structural characterisation of neutrophil glycans by ultra sensitive mass spectrometric glycomics methodology Structural characterization of non-reducing oligosaccharide produced by Arthrobacter crystallopoietes N-08 Fragmentation of deprotonated D-ribose and D-fructose in MALDI-Comparison with dissociative electron attachment Multidimensional profiling of components in complex mixtures of natural products for metabolic analysis, proof of concept: Application to Quillaja saponins Structural investigation of bacterial lipopolysaccharides by mass spectrometry and tandem mass spectrometry Probe design and synthesis of Galb(1 ! 3)[NeuAca(2 ! 6)]GlcNAcb(1 ! 2)Man motif of N-glycan Glycoprofiling bifidobacterial consumption of galacto-oligosaccharides by mass spectrometry reveals strain-specific, preferential consumption of glycans N-Glycosylation of plant recombinant pharmaceuticals Synthesis of thiourea-tethered Cglycosyl amino acids via isothiocyanate-amine coupling Permeate from cheese whey ultrafiltration is a source of milk oligosaccharides Detecting glycan cancer biomarkers in serum samples using MALDI FT-ICR mass spectrometry data Quantitative analysis of glycation sites on human serum albumin using 16 O/ 18 Olabeling and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry Characterization of WRSs2 and WRSs3, new second-generation virG(icsA)-based Shigella sonnei vaccine candidates with the potential for reduced reactogenicity Mass spectrometry in the characterization of human genetic N-glycosylation defects Combined treatment of human MCF-7 breast carcinoma with antibody, cationic lipid and hyaluronic acid using ex vivo assays Ex vivo assays of CEM cells cultured and treated in the three dimensional cultures Glycan analysis and influenza A virus infection of primary swine respiratory epithelial cells: The importance of NeuAca2-6 glycans Acceptor substrate discrimination in phosphatidyl-myo-inositol mannoside synthesis. Structural and mutational analysis of mannosyl transferase Corynebacterium glutamicum PimB Synthesis and antimicrobial evaluation of amphiphilic neamine derivatives Clicking pentafluorostyrene copolymers: Synthesis, nanoprecipitation, and glycosylation Trends in glycosylation, glycoanalysis and glycoengineering of therapeutic antibodies and Fc-fusion proteins Chemical Structure of Bacteriovorax stolpii Lipid A Core chirality based tailoring of the liquid crystalline properties of supermolecular tetrapedes Development of a nanoLC LTQ orbitrap mass spectrometric method for profiling glycans derived from plasma from healthy, benign tumor control, and epithelial ovarian cancer patients Development of a robust and high throughput method for profiling N-linked glycans derived from plasma glycoproteins by nanoLC-FTICR mass spectrometry Genomic and biochemical analysis of N-glycosylation in the mushroom-forming basidiomycete Schizophyllum commune The structurally similar, penta-acylated lipopolysaccharides of Porphyromonas gingivalis and Bacteroides elicit strikingly different innate immune responses Design, synthesis and biological evaluation of carbohydrate-functionalized cyclodextrins and liposomes for hepatocyte-specific targeting Ex post glycoconjugation of phthalocyanines Modification of recombinant proteins by covalent polysialation illustrated with the example of human insulin Influence of protein molecular mass on the glycation Historical overview of glycoanalysis Synthesis of differentially protected glucosamine building blocks and their evaluation as glycosylating agents De novo synthesis of differentially protected L-iduronic acid glycosylating agents Chemical analysis of flavonoid constituents of the seagrass Halophila stipulacea: First finding of malonylated derivatives in marine phanerogams Substantial spatial flexibility and hydrogen bonding within the catalysis exerted by cyclodextrin artificial glycosidases Lysine and arginine side chains in glycosaminoglycanprotein complexes investigated by NMR, cross-linking, and mass spectrometry: A case study of the factor H-heparin interaction Glycomic analysis of N-linked carbohydrate epitopes from CD24 of mouse brain O-Glycosylation pattern of CD24 from mouse brain A high-throughput Oglycopeptide discovery platform for seromic profiling A high-throughput Oglycopeptide discovery platform for seromic profiling-Correction Alteration of protein glycosylation in liver diseases Investigating the molecular basis for the virulence of Escherichia coli K5 by nuclear magnetic resonance analysis of the capsule polysaccharide Monitoring of malting process by characterization of glycation of barley protein Z Determination of glycan structure from tandem mass spectra Use of fullerene-, octadecyl-, and triaconthyl silica for solid phase extraction of tryptic peptides obtained from unmodified and in vitro glycated human serum albumin and fibrinogen Site specific conjugation of fluoroprobes to the remodeled Fc N-glycans of monoclonal antibodies using mutant glycosyltransferases: Application for cell surface antigen detection Innovative approach for producing injectable, biodegradable materials using chitooligosaccharides and green chemistry Reciprocal principle of molecular recognition in supramolecular chromatography-highly selective analytical separation of cyclodextrin congeners on a silica-bonded [60]fullerene stationary phase Cation-independent mannose 6-phosphate receptor. A composite of distinct phosphomannosyl binding sites Synthesis of LacdiNAc-terminated glycoconjugates by mutant galactosyltransferase-A way to new glycodrugs and materials New 4,4-difluoro-3a,4a-diaza-sindacene (BODIPY)-labeled sphingolipids for membrane studies Solid-phase synthesis of a pentavalent GalNAc-containing glycopeptide (Tn antigen) representing the nephropathy-associated IgA hinge region Chiral-auxiliary-mediated 1,2-cis-glycosylations for the solid-supported synthesis of a biologically important branched a-glucan Multimeric bivalent immunogens from recombinant tetanus toxin HC fragment, synthetic hexasaccharides, and a glycopeptide adjuvant The elucidation of the structure of Thermotoga maritima peptidoglycan reveals two novel types of cross-link A M23B family metallopeptidase of Helicobacter pylori required for cell shape, pole formation and virulence Lysosomal storage of oligosaccharide and glycosphingolipid in imino sugar treated cells Multivalent dendrimeric compounds containing carbohydrates expressed on immune cells inhibit infection by primary isolates of HIV-1 Ion exchange and purification of carbohydrates on a Nafion(R) membrane as a new sample pretreatment for matrix-assisted laser desorption-ionization mass spectrometry Chinese hamster ovary cells can produce galactose-a-1,3-galactose antigens on proteins Characterization of AtlL, a bifunctional autolysin of Staphylococcus lugdunensis with Nacetylglucosaminidase and N-acetylmuramoyl-L-alanine amidase activities Dimeric architecture of the Hendra virus attachment glycoprotein: Evidence for a conserved mode of assembly Unusual molecular architecture of the Machupo virus attachment glycoprotein Synthesis of polymerizable cyclodextrin derivatives for use in adhesion-promoting monomer formulations Physicochemical properties of microbial glycopolymers An analytical system for the characterization of highly heterogeneous mixtures of N-linked oligosaccharides Site directed processing: Role of amino acid sequences and glycosylation of acceptor glycopeptides in the assembly of extended mucin type O-glycan core 2 High content phenotypic cell-based visual screen identifies Mycobacterium tuberculosis acyltrehalose-containing glycolipids involved in phagosome remodeling Strategies for analysis of the glycosylation of proteins: Current status and future perspectives Characterization of IRX10 and IRX10-like reveals an essential role in glucuronoxylan biosynthesis in Arabidopsis UV-MALDI-TOF mass spectrometry analysis of heparin oligosaccharides obtained by nitrous acid controlled degradation and high performance anion exchange chromatography Distribution of O-glycosylhydrolases in marine fungi of the Sea of Japan and the Sea of Okhotsk: Characterization of exocellular N-acetyl-b-D-glucosaminidase of the marine fungus Penicillium canescens Derivatization in mass spectrometry Identification, characterization, and biosynthesis of a novel N-glycan modification in the fruiting body of the basidiomycete Coprinopsis cinerea Lipid remodelling of glycosylphosphatidylinositol (GPI) glycoconjugates in procyclic form trypanosomes: Biosynthesis and processing of GPIs revisited Caenorhabditis elegans N-glycan core b-galactoside confers sensitivity towards nematotoxic fungal galectin CGL2 Novel mannosidase inhibitors probe glycoprotein degradation pathways in cells Chitosan oligosaccharides modulate the supramolecular conformation and the biological activity of oligogalacturonides in Arabidopsis Efficient synthesis of a 6-deoxytalose tetrasaccharide related to the antigenic O-polysaccharide produced by Aggregatibacter actinomycetemcomitans serotype c Aniline/a-cyano-4-hydroxycinnamic acid is a highly versatile ionic liquid for matrix-assisted laser desorption/ionization mass spectrometry 1H-Pteridine-2,4-dione (lumazine): A new MALDI matrix for complex (phospho)lipid mixtures analysis Characterization of Acp, a peptidoglycan hydrolase of Clostridium perfringens with Nacetylglucosaminidase activity that is implicated in cell separation and stress-induced autolysis GlycoBase and autoGU: Tools for HPLC-based glycan analysis Carrageenans: Biological properties, chemical modifications and structural analysis-A review Click" glycodendrimers containing 27, 81 and 243 modified xylopyranoside termini The plasma von Willebrand factor O-glycome comprises a surprising variety of structures including ABH antigens and disialosyl motifs Analytical progress for protein glycosylation in China Strategies for proteomic analysis of nonenzymatically glycated proteins Preparation of saturated and unsaturated fatty acid hydrazides and long chain C-glycoside ketohydrazones Glycoproteome study in myocardial lesions serum by integrated mass spectrometry approach: Preliminary insights Changes of serum-associated fucosylated glycoproteins and changes in glycosylation of IgA in human cirrhosis Efficient isolation of the subunits of recombinant and pituitary glycoprotein hormones Ferrocene-cyclodextrin conjugates: Synthesis, supramolecular behavior, and use as electrochemical sensors Ferrocene-carbohydrate conjugates as electrochemical probes for molecular recognition studies Characterization of main anthocyanins extracted from pericarp blue corn by MALDI-ToF MS Highly glycosylated human alpha interferon: An insight into a new therapeutic candidate The protease-associated domain and C-terminal extension are required for zymogen processing, sorting within the secretory pathway, and activity of tomato subtilase 3 (SlSBT3) Design and creativity in synthesis of multivalent neoglycoconjugates 3-Deoxy-Dmanno-octulosonic acid (Kdo) hydrolase identified in Francisella tularensis, Helicobacter pylori, and Legionella pneumophila Synthesis and characterization of biotin chainend functionalized boronic acid-containing polymer (boropolymer) as functional glyco-affinity macroligand MALDI mass spectrometry imaging of gangliosides in mouse brain using ionic liquid matrix DAS181 Inhibits H5N1 influenza virus infection of human lung tissues Synthesis, characterization, and self-assembled nanofibers of carbohydrate-functionalized mono-and di(2,2 0 :6 0 ,2 00 -terpyridinyl)arenes Developmental regulation of oligosialylation in zebrafish Glycan array on aluminum oxide-coated glass slides through phosphonate chemistry Mannose receptor interacts with Fc receptors and is critical for the development of crescentic glomerulonephritis in mice Efficient synthesis of Idraparinux, the anticoagulant pentasaccharide The efficient total synthesis of bisglycosyl apigenin from naringenin: A greener way A mixed cyclodextrinbiphenyl thermotropic liquid crystal: Synthesis, liquid-crystalline properties, and supramolecular organization Boronate affinity monolith for highly selective enrichment of glycopeptides and glycoproteins Application of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) in preparation of chitosan oligosaccharides (COS) with degree of polymerization (DP) 5-12 containing well-distributed acetyl groups Study of on-resin convergent synthesis of N-linked glycopeptides containing a large high mannose N-linked oligosaccharide Facile synthesis of cyclopeptide-centered multivalent glycoclusters with 'click chemistry' and molecular recognition study by surface plasmon resonance Glycosite analysis in glycoproteomics by mass spectrometry Combination of matrix-assisted laser desorption ionization and electrospray ionization mass spectrometry for the analysis of intact glycopeptides from horseradish peroxidase An introduction to sphingolipid metabolism and analysis by new technologies One-pipeline approach achieving glycoprotein identification and obtaining intact glycopeptide information by tandem mass spectrometry Imaging MALDI mass spectrometry of sphingolipids using an oscillating capillary nebulizer matrix application system Single-crystalline EuF3 hollow hexagonal microdisks: Synthesis and application as a background-free matrix for MALDI-TOF-MS analysis of small molecules and polyethylene glycols Selective desorption/ ionization of sulfatides by MALDI-MS facilitated using 9-aminoacridine as matrix Tethered derivatives of D-glucose and pentacyclic triterpenes for homo/heterobivalent inhibition of glycogen phosphorylase Total synthesis of a furostan saponin, timosaponin BII Subcompartment localization of the side chain xyloglucan-synthesizing enzymes within Golgi stacks of tobacco suspension-cultured cells Carbohydrates as recognition receptors in biosensing applications Expression system for human glycosyltransferases and its application Production, purification, and characterization of human a1 proteinase inhibitor from Aspergillus niger Direct amidation of aldoses and decarboxylative amidation of a-keto acids: An efficient conjugation method for unprotected carbohydrate molecules Novel succinylated and large-sized osmoregulated periplasmic glucans of Pseudomonas syringae pv. syringae Chiral separation of hesperetin and hesperetin-O-glycoside in capillary electrophoresis using microbial b-1,2-glucans Structural modification and characterization of rice starch treated by Thermus aquaticus 4-a-glucanotransferase Enzymatic biosynthesis of a puerarincycloamylose inclusion complex by 4-aglucanotransferase and maltogenic amylase Analysis of mixture of maltooligoses using MALDI-TOFMS: Influence of cationizing agent types Comparison of ionization behaviors of ring and linear carbohydrates in MALDI-TOFMS Challenges of determining O-glycopeptide heterogeneity: A fungal glucanase model system Mutational analysis of Bacillus megaterium QM B1551 cortex-lytic enzymes Mass spectrometric imaging for biomedical tissue analysis Synthesis of MUC1 peptide and glycopeptide dendrimers Synthesis and biological evaluation of multivalent carbohydrate ligands obtained by click assembly of pseudo-rotaxanes Pseudomonas aeruginosa exploits lipid A and muropeptides modification as a strategy to lower innate immunity during cystic fibrosis lung infection Straightforward synthesis of novel Akt inhibitors based on a glucose scaffold The role of sugar configuration in the acetolysis of 6-deoxyhexose methyl glycosides Synthesis of a b-GlcN-(1 ! 4)-MurNAc building block en route to N-deacetylated peptidoglycan fragments A urea-linked glucosamine dimer as a building block for the synthesis of linear and cyclic neosaccharides A simple and rapid method for the permethylation of carbohydrates Trypanosoma brucei AMP-activated kinase subunit homologs influence surface molecule expression Porphyromonas gingivalis resistance to polymyxin B is determined by the lipid A 4 0 -phosphatase, PGN_0524 A new archaeal b-glycosidase from Sulfolobus solfataricus. Seeding a novel retaining b-glycan-specific glycoside hydrolase family along with the human non-lysosomal glucosylceramidase CBA2 b-Glycosyl azides as substrates for a-glycosynthases: Preparation of efficient a-L-fucosynthases Synthetic heparan sulfate oligosaccharides Inhibit endothelial cell functions essential for angiogenesis Towards the synthesis of 1-deoxy-1-nitropiperidinoses Synthesis and evaluation of S-and C(1)-substituted analogues of lincomycin Localization and imaging of sialylated glycosphingolipids in brain tissue sections by MALDI mass spectrometry An efficient modular approach for the assembly of S-linked glycopeptoids Glycosidase inhibition with fullerene iminosugar balls: A dramatic multivalent effect Modification of gastric mucin oligosaccharide expression in rhesus macaques after infection with Helicobacter pylori Identification of novel contributions to high-affinity glycoprotein-receptor interactions using engineered ligands Synthesis and conformational analysis of a novel carbohydrate-fused bis-crown ether: Crown-CyPLOS Design, synthesis and characterisation of a fluorescently labelled CyPLOS ionophore Application of liquid chromatography-tandem mass spectrometry for the characterization of galactosylated and tagatosylated b-lactoglobulin peptides derived from in vitro gastrointestinal digestion Effect of glycation on the gastrointestinal digestibility and immunoreactivity of bovine b-lactoglobulin Role of pyridoxamine in the formation of the Amadori/Heyns compounds and aggregates during the glycation of b-lactoglobulin with galactose and tagatose Acceptor products of alternansucrase with gentiobiose. Production of novel oligosaccharides for food and feed and elimination of bitterness Glucosylation of raffinose via alternansucrase acceptor reactions Investigating the candidacy of LPS-based glycoconjugates to prevent invasive meningococcal disease: Chemical strategies to prepare glycoconjugates with good carbohydrate loading Investigating the candidacy of LPS-based glycoconjugates to prevent invasive meningococcal disease: Immunology of glycoconjugates with high carbohydrate loading Towards a second generation of ionic liquid matrices (ILMs) for MALDI-MS of peptides, proteins, and carbohydrates Sinorhizobium fredii HH103 cgs mutants are unable to nodulate determinate-and indeterminate nodule-forming legumes and overproduce an altered EPS Carbohydrate and domain architecture of an immature antibody glycoform exhibiting enhanced effector functions A human embryonic kidney 293T cell line mutated at the Golgi a-mannosidase II locus Sieve-based device for MALDI sample preparation. II. Instrumental parameterization A link between the assembly of flagella and lipooligosaccharide of the Gram-negative bacterium Campylobacter jejuni Nonglycosidically linked pseudodisaccharides: Thioethers, sulfoxides, sulfones, ethers, selenoethers, and their binding to lectins Stable expression of a human-like sialylated recombinant thyrotropin in a Chinese hamster ovary cell line expressing a2,6-sialyltransferase Impaired lysosomal trimming of N-linked oligosaccharides leads to hyperglycosylation of native lysosomal proteins in mice with a-mannosidosis Biochemical and immunocytological characterizations of Arabidopsis thaliana pollen tube cell wall Engineering of N. benthamiana L. plants for production of N-acetylgalactosamine-glycosylated proteins-Towards development of a plant-based platform for production of protein therapeutics with mucin type O-glycosylation Xylan-based nanoparticles: Prodrugs for ibuprofen release Towards unnatural xylan based polysaccharides: Reductive amination as a tool to access highly engineered carbohydrates A simple micromethod for determining precise oligosaccharidic specificity of mannose-binding lectins Functional and morphological adaptation to peptidoglycan precursor alteration in Lactococcus lactis Towards the implementation of quality by design to the production of therapeutic monoclonal antibodies with desired glycosylation patterns Molecular characterization of a putative sucrose: Fructan 6-fructosyltransferase (6-SFT) of the cold-resistant Patagonian grass Bromus pictus associated with fructan accumulation under low temperatures Biomarkers and diagnosis of congenital disorders of glycosylation Improved procedures for the selective chemical fragmentation of rhamnogalacturonans When can glycopeptides be assigned based solely on high-resolution mass spectrometry data? MmpS4 promotes glycopeptidolipids biosynthesis and export in Mycobacterium smegmatis Glyco-Spectrum-Scan: Fishing glycopeptides from MS spectra of protease digests of human colostrum sIgA Common sialylated glycan in Actinobacillus suis The major surface carbohydrates of the Echinococcus granulosus cyst: Mucin-type O-glycans decorated by novel galactosebased structures Surface analytical characterization of carbohydrate microarrays Shiga toxin receptor Gb3Cer/ CD77: Tumor-association and promising therapeutic target in pancreas and colon cancer Synthesis of some quaternary N-(1,4-anhydro-5-deoxy-D,L-ribitol-5-yl)ammonium salts A barley cellulose synthase-like CSLH gene mediates (1,3;1,4)-b-D-glucan synthesis in transgenic Arabidopsis Analytical performance of immobilized pronase for glycopeptide footprinting and implications for surpassing reductionist glycoproteomics Recombinant expression and characterization of N-acetylglucosaminyltransferase I derived from Nicotiana tabacum Comparison of the N-linked glycosylation of human b1,3-N-acetylglucosaminyltransferase 2 expressed in insect cells and silkworm larvae Improved secretion of molecular chaperone-assisted human IgG in silkworm, and no alterations in their N-linked glycan structures Structure of glycosylated Cu/Znsuperoxide dismutase from kluyveromyces yeast NBIMCC 1984 Glycan structures and antiviral effect of the structural subunit RvH2 of Rapana hemocyanin BacA, an ABC transporter involved in maintenance of chronic murine infections with Mycobacterium tuberculosis A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates Transformation of linear oligoketosides into macrocyclic neoglycoconjugates A new ligation strategy for peptide and protein glycosylation: Photoinduced thiol-ene coupling Graphene as a novel matrix for the analysis of small molecules by MALDI-TOF MS Envelope glycans of immunodeficiency virions are almost entirely oligomannose antigens Clinical collection and protein properties of expressed prostatic secretions as a source for biomarkers of prostatic disease Mycobacterium marinum MMAR-2380, a predicted transmembrane acyltransferase, is essential for the presence of the mannose cap on lipoarabinomannan Electrochemically controlled adsorption of Fc-functionalized polymers on b-CD-modified selfassembled monolayers Production of carbohydrate building blocks from red seaweed polysaccharides. Efficient conversion of galactans into C-glycosyl aldehydes Polysaccharide mimicry of the epitope of the broadly neutralizing anti-HIV antibody, 2G12, induces enhanced antibody responses to self oligomannose glycans Glycosylated cell-penetrating peptides and their conjugates to a proapoptotic peptide: Preparation by click chemistry and cell viability studies 355 nm Multiphoton dissociation and ionization of 2,5-dihydroxyacetophenone Lack of complex N-glycans on HIV-1 envelope glycoproteins preserves protein conformation and entry function Prebiotic oligosaccharides: In vitro evidence for gastrointestinal epithelial transfer and immunomodulatory properties C-Furyl glycosides, II: Synthesis and antimicrobial evaluation of C-furyl glycosides bearing pyrazolines, isoxazolines, and 5,6-dihydropyrimidine-2(1H)-thiones Structural characterization of Bordetella parapertussis lipid A Polysaccharide pharmacokinetics: Amphotericin B arabinogalactan conjugate -a drug delivery system or a new pharmaceutical entity Hexosamine template. A platform for modulating gene expression and for sugar-based drug discovery Combinatorial syntheses of trisaccharide libraries on a soluble polymeric support Lipophilic thioglycosides for the solution-phase synthesis of oligosaccharides using biphasic liquid-liquid separation Chiral and structural analysis of biomolecules using mass spectrometry and ion mobility-mass spectrometry Fluorescent labeling of a carboxyl group of sialic acid for MALDI-MS analysis of sialyloligosaccharides and ganglioside Liquid chromatography combined with mass spectrometry for the investigation of endoglucanase selectivity on carboxymethyl cellulose Investigation of endoglucanase selectivity on carboxymethyl cellulose by mass spectrometric techniques Cellulosic graft copolymer: Poly(methyl methacrylate) with cellulose side chains Radially oriented cellulose triacetate chains on gold nanoparticles Amphiphilic carbohydrate-phthalocyanine conjugates obtained by glycosylation or by azide-alkyne click reaction Different and new Nod factors produced by Rhizobium tropici CIAT899 following Na þ stress Identification of the polyketide synthase involved in the biosynthesis of the surface-exposed lipooligosaccharides in mycobacteria Bioanalysis of recombinant proteins and antibodies by mass spectrometry Lectin and carbohydrate microarrays: New high-throughput methods for glycoprotein, carbohydrate-binding protein and carbohydrate-active enzyme analysis Chitosans as bioactive macromolecules to protect economically relevant crops from their main pathogens Serum N-glycome biomarker for monitoring development of DENA-induced hepatocellular carcinoma in rat Secondary cell wall formation in Cryptococcus neoformans as a rescue mechanism against acid-induced autolysis Identification and quantification of protein posttranslational modifications Matrixassisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry analysis of oligosaccharides and oligosaccharide alditols obtained by hydrolysis of agaroses and carrageenans, two important types of red seaweed polysaccharides Glycopeptidome of a heavily N-glycosylated cell surface glycoprotein of Dictyostelium implicated in cell adhesion Fragmentation behavior of Amadori-peptides obtained by non-enzymatic glycosylation of lysine residues with ADP-ribose in tandem mass spectrometry Effect of 1,3;1,6-b-D-glucan and the products of its enzymatic transformation on the formation of germs of buckwheat Fagopyrum esculentum Mönch Synthesis of 3,6-branched arabinogalactan-type tetra-and hexasaccharides for characterization of monoclonal antibodies Phosphoglucomutase of Yersinia pestis is required for autoaggregation and polymyxin B resistance Mass spectrometric investigation of molecular variability of grass pollen group 1 allergens Cyclodextrin aldehydes are oxidase mimics Simultaneous glycoproteomics on the basis of structure using ion mobility-mass spectrometry Characterizing ion mobility-mass spectrometry conformation space for the analysis of complex biological samples Effect of ionic liquids as additives in the catalytic properties of different immobilized preparations of Rhizomucor miehei lipase in the hydrolysis of peracetylated lactal Synthetic pseudopterosin analogues: A novel class of antiinflammatory drug candidates Purification, characterization and in vivo studies of salmon heparin Thieme chemistry journal awardees-Where are they now? Synthesis of the marine glycolipid dioctadecanoyl discoside Thiyl glycosylation of olefinic proteins: S-linked glycoconjugate synthesis Synthesis of the Lewis b pentasaccharide and a HSA-conjugate thereof MALDI mass spectrometry imaging, from its origins up to today: The state of the art Detection of honeybee venom in envenomed tissues by direct MALDI MSI Bioinformatics and molecular modeling in glycobiology Expression of rat b(1,4)-N-acetylglucosaminyltransferase III in Nicotiana tabacum remodels the plantspecific N-glycosylation Glycoprotein expression in human milk during lactation 1-Deoxynojirimycins with dansyl capped N-substituents as probes for Morbus Gaucher affected cell lines New secoiridoid glucosides from Ligustrum lucidum induce ERK and CREB phosphorylation in cultured cortical neurons Application of MALDI-TOF mass spectrometry in lipidomics An update of MALDI-TOF mass spectrometry in lipid research MALDI-TOF-MS directly combined with TLC: A review of the current state Isolation and structural analysis in vivo of newly synthesized fructooligosaccharides in onion bulbs tissues (Allium cepa L.) during storage MALDI mass spectrometry using 2,4,6-trihydroxyacetophenone and 2,4-dihydroxyacetophenone with cyclodextrins: Suppression of matrix-related ions in low-molecular-weight region Change in glycosylation pattern with extension of endoplasmic reticulum retention signal sequence of mouse antibody produced by suspensioncultured tobacco BY2 cells Chemical characterization of the oligosaccharides in Bactrian camel (Camelus bactrianus) milk and colostrum Dendritic sugar-microarrays by click chemistry Aggregation of Alzheimer amyloid b peptide (1-42) on the multivalent sulfonated sugar interface Scope and limitations of imidazolium-based ionic liquids as room temperature glycosylation promoters The carboxy-terminal ERretention motif, SEKDEL, influences the N-linked glycosylation of recombinant human a-L-iduronidase but has little effect on enzyme activity in seeds of Brassica napus and Nicotiana tabacum Mass spectrometric fragmentation analysis of oligosialic and polysialic acids Quantification of nucleotide-activated sialic acids by a combination of reduction and fluorescent labeling Synaptic cell adhesion molecule SynCAM 1 is a target for polysialylation in postnatal mouse brain Comparative characterization of the Arabidopsis subfamily a1 b-galactosidases Preparation and gelling properties of sugar-contained low-molecular-mass gelators: Combination of cholesterol and linear glucose Generation of asparagine-linked glycan structure databases and their use The first total synthesis of 7-O-b-Dglucopyranosyl-4 0 -O-a-L-rhamnopyranosyl apigenin via a hexanoyl ester-based protection strategy Efficient synthesis of 4-amido-N 5 -acetyl-4-deoxyneuraminic acid and its application to the C-4 modification of sialic acids Endothelial galectin-1 binds to specific glycans on Nipah virus fusion protein and inhibits maturation, mobility, and function to block syncytia formation Lectin-directed enzyme activated prodrug therapy (LEAPT): Synthesis and evaluation of rhamnosecapped prodrugs Characterization of a new b(1-3)-glucan branching activity of Aspergillus fumigatus Enhanced production and partial characterization of an extracellular polysaccharide from newly isolated Azotobacter sp. SSB81 Phase diagrams of monoacylated amide-linked disaccharide glycolipids Diamond, titanium dioxide, titanium silicon oxide, and barium strontium titanium oxide nanoparticles as matrixes for direct matrix-assisted laser desorption/ionization mass spectrometry analysis of carbohydrates in plant tissues Synthesis of a tetrasaccharide corresponding to the teichoic acid from the cell wall of Streptomyces sp. VKM Ac-2275 Synthesis of the hexasaccharide repeating unit corresponding to the cell wall lipopolysaccharide of Azospirillum irakense KBC1 Sialic acids in different Leishmania sp., its correlation with nitric oxide resistance and host responses 9-O-acetylated sialic acids enhance entry of virulent Leishmania donovani promastigotes into macrophages High throughput quantification of N-glycans using one-pot sialic acid modification and matrix assisted laser desorption ionization time-offlight mass spectrometry GDP-D-mannose 3,5-epimerase (GME) plays a key role at the intersection of ascorbate and non-cellulosic cell-wall biosynthesis in tomato An HPLC-MALDI MS method for N-glycan analyses using smaller size samples: Application to monitor glycan modulation by medium conditions Towards a reliable molecular mass determination of intact glycoproteins by matrixassisted laser desorption/ionization time-of-flight mass spectrometry Ionic liquid matrices for MALDI-TOF-MS analysis of intact glycoproteins Synthesis of PEGylated lactose analogs for inhibition studies on T. cruzi trans-sialidase Glycosylation site-specific analysis of clade C HIV-1 envelope proteins Enzymatic/chemical release of O-glycans allowing MS analysis at high sensitivity Glycomic profiling of invasive and non-invasive breast cancer cells Assessment of chemoselective neoglycosylation methods using chlorambucil as a model Glycan family analysis for deducing N-glycan topology from single MS Detection of hepatocellular carcinoma using glycomic analysis Discovery of rifampicin as a new anti-glycating compound by matrix-assisted laser desorption/ionization mass spectrometry-based insulin glycation assay First total synthesis of 1,2-dipalmitoyl-3-(N-palmitoyl-6 0 -amino-6 0 -deoxy-a-Dglucosyl)-sn-glycerol-a glycoglycerolipid of a marine alga with a high inhibitor activity against human Myt1-kinase Acidiplasma aeolicum gen. nov., sp. nov., a euryarchaeon of the family Ferroplasmaceae isolated from a hydrothermal pool, and transfer of Ferroplasma cupricumulans to Acidiplasma cupricumulans comb. nov Synthesis of polyhydroxy [n]-polyurethanes derived from a carbohydrate precursor Characterization of N-linked glycosylation on recombinant glycoproteins produced in Pichia pastoris using ESI-MS and MALDI-TOF DNA-templated homo-and heterodimerization of peptide nucleic acid encoded oligosaccharides that mimick the carbohydrate epitope of HIV Human macrophage activation triggered by chitotriosidase-mediated chitin and chitosan degradation Evaluation of conditions for release of mucin-type oligosaccharides from glycoproteins by hydrazine gas treatment Imaging mass spectrometry of glycolipids Visualization of spatial distribution of g-aminobutyric acid in eggplant (Solanum melongena) by matrix-assisted laser desorption/ionization imaging mass spectrometry The specific localization of seminolipid molecular species on mouse testis during testicular maturation revealed by imaging mass spectrometry The detection of glycosphingolipids in brain tissue sections by imaging mass spectrometry using gold nanoparticles Practical heavy fluorous tag for carbohydrate synthesis with minimal chromatographic purification Production and Nglycosylation of recombinant human cell adhesion molecule L1 from insect cells using the stable expression system. Effect of dimethyl sulfoxide Glyco-SAMs by 'dual click': Thiourea-bridged glyco-OEG azides for cycloaddition on surfaces Mass spectrometry in the elucidation of the glycoproteome of bacterial pathogens Investigation towards bivalent chemically defined glycoconjugate immunogens prepared from acid-detoxified lipopolysaccharide of Vibrio cholerae O1, serotype Inaba The synthesis and immune stimulating action of mannose-capped lysine-based dendrimers Some patterns in dimer II formation in BODIPY-FL-labeled lipids BODIPY-labeled ganglioside probes for membrane and biological studies Development of a high performance anion exchange chromatography analysis for mapping of oligosaccharides Analytical approaches towards the structural characterization of microbial wall glycopolymers Differential effect of plasma or erythrocyte AGE-ligands of RAGE on expression of transcripts for receptor isoforms Biosynthesis of a new UDPsugar, UDP-2-acetamido-2-deoxyxylose, in the human pathogen Bacillus cereus subspecies cytotoxis NVH 391-98 Effective enlargement of fluorescence resonance energy transfer of poly-porphyrin mediated by b-cyclodextrin dimers Total synthesis of the heptasaccharide repeating unit of the iron-binding exopolysaccharide secreted by Klebsiella oxytoca BAS-10 Mass spectrometry characterization of the glycation sites of bovine insulin by tandem mass spectrometry Oxidative modifications in glycated insulin New insights into the early steps of phosphatidylinositol mannoside biosynthesis in mycobacteria. PimB' is an essential enzyme of Mycobacterium smegmatis Fatty acyl structures of Mycobacterium tuberculosis sulfoglycolipid govern T cell response Automated measurement of permethylated serum N-glycans by MALDI-linear ion trap mass spectrometry Monoterpenoid glucoindole alkaloids and iridoids from Pterocephalus pinardii Production, refining, structural characterization and fermentability of rice husk xylooligosaccharides OLIgo mass profiling (OLIMP) of extracellular polysaccharides Facile synthesis of three bidesmosidic oleanolic acid saponins with strong inhibitory activity on pancreatic lipase Sortase-catalyzed peptide-Glycosylphosphatidylinositol analogue ligation One-pot synthesis of cyclic antifreeze glycopeptides Syntheses of glycoclusters containing a phosphocholine residue related to a glycosphingolipid from the earthworm Pheretima hilgendorfi Establishment of a real-time analytical method for free oligosaccharide transport from the ER to the cytosol Purification, characterization and molecular cloning of a novel endo-b-N-acetylglucosaminidase from the basidiomycete, Flammulina velutipes Comparative glycomic profiling in esophageal adenocarcinoma Derivatization and analysis of oligosaccharides by matrix-assisted laser desorption/ionization time of-flight mass spectrometry Sulfated oligosaccharide cluster with polylysine core scaffold as a new anti-HIV dendrimer Lipophilic b-cyclodextrin cyclic-nitrone conjugate: synthesis and spin trapping studies Sialylation using N-glycolylneuraminyl phosphite donors to synthesize Neu5Gc-containing glycans Chemical de-Oglycosylation of glycoproteins for application in LC-based proteomics Secondary acylation of Vibrio cholerae lipopolysaccharide requires phosphorylation of Kdo Rapid characterization of N-linked glycans from secreted and gel-purified monoclonal antibodies using MALDI-ToF mass spectrometry Synthesis of porphyrin-carbohydrate conjugates using "click" chemistry and their preliminary evaluation in human HEp2 cells An improved protocol for N-glycosylation analysis of gel-separated sialylated glycoproteins by MALDI-TOF/ TOF Artificial polymerases and molecular chaperones Interaction of Nectarin 4 with a fungal protein triggers a microbial surveillance and defense mechanism in nectar N-Glycans on the link domain of human HARE/Stabilin-2 are needed for hyaluronan binding to purified ecto-domain, but not for cellular endocytosis of hyaluronan Analytical characterization of monoclonal antibodies: Linking structure to function Identification of Rhodococcus equi lipids recognized by host cytotoxic T lymphocytes Matrix-assisted laser desorption/ionization mass spectrometry of carbohydrates Analysis of carbohydrates and glycoconjugates by matrixassisted laser desorption/ionization mass spectrometry: An update covering the period 1999-2000 Analysis of carbohydrates and glycoconjugates by matrixassisted laser desorption/ionization mass spectrometry: An update covering the period Analysis of carbohydrates and glycoconjugates by matrixassisted laser desorption/ionization mass spectrometry: An update for Electrospray and MALDI mass spectrometry: Fundamentals, instrumentation, practicalities, and biological applications Analysis of carbohydrates and glycoconjugates by matrixassisted laser desorption/ionization mass spectrometry: An update for the period Analysis of carbohydrates and glycoconjugates by matrixassisted laser desorption/ionization mass spectrometry: An update for Identification of by-products formed during the release of N-glycans with protein N-glycosidase F in the presence of dithiothreitol Application of negative ion MS/ MS to the identification of N-glycans released from carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1) Structural and quantitative analysis of N-linked glycans by MALDI and negative ion nanospray mass spectrometry Proposal for a standard system for drawing structural diagrams of N-and O-linked carbohydrates and related compounds Fragmentation of negative ions from N-linked carbohydrates, Part 4. Fragmentation of complex glycans lacking substitution on the 6-antenna Identification of high-mannose and multiantennary complextype N-linked glycans containing a-galactose epitopes from Nurse shark IgM heavy chain O-Glycoside biomarker of apolipoprotein C3: Responsiveness to obesity, bariatric surgery, and therapy with metformin, to chronic or severe liver disease and to mortality in severe sepsis and graft vs host disease Pyridylamination as a means of analyzing complex sugar chains Switching the selectivity of a polyglycerol dendrimer monomolecularly imprinted with D-(À)-fructose KEGG GLYCAN for integrated analysis of pathways, genes and glycan structures Sphingolipidomics: Methods for the comprehensive analysis of sphingolipids Antifreeze glycopeptide analogues: Microwaveenhanced synthesis and functional studies Degradation of chitosans with a family 46 chitosanase from Streptomyces coelicolor A3(2) Glycomics of bone marrow-derived mesenchymal stem cells can be used to evaluate their cellular differentiation stage Rational engineering of mannosyl binding in the distal glycone subsites of Cellulomonas fimi Endo-b-1,4-mannanase: Mannosyl binding promoted at subsite -2 and demoted at subsite -3 UV-laser ablation of ionic liquid matrices Structure analysis of N-glycoproteins Toward synthesis of the isosteric sulfonate analogues of the AT-III binding domain of heparin Plant immunity induced by oligogalacturonides alters root growth in a process involving flavonoid accumulation in Arabidopsis thaliana Conversion of a-amyrin into centellosides by plant cell cultures of Centella asiatica Carbohydrate analysis throughout the development of a protein therapeutic Positive and negative analyte ion yield in matrix-assisted laser desorption/ionization revisited g-Cyclodextrins possessing an azido group and a triisopropylbenzenesulfonyl group as useful synthetic and authentic intermediates for unsymmetrically functionalized derivatives Hemicellulase production in Chrysosporium lucknowense C1 Genetically engineered mannosylated-human serum albumin as a versatile carrier for liverselective therapeutics Free oligosaccharides to monitor glycoprotein endoplasmic reticulumassociated degradation in Saccharomyces cerevisiae Chemical synthesis, folding, and structural insights into O-fucosylated epidermal growth factor-like repeat 12 of mouse notch-1 receptor A water-soluble ruthenium glycosylated porphyrin catalyst for carbenoid transfer reactions in aqueous media with applications in bioconjugation reactions Synthesis and self-assembled nanostructures of novel chiral amphiphilic liquid crystals containing b-D-galactopyranoside end-groups Synthetic antitumor vaccines from tetanus toxoid conjugates of MUC1 glycopeptides with the Thomsen-Friedenreich antigen and a fluorine-substituted analogue Matrix-free UV-laser desorption/ionization (LDI) mass spectrometric imaging at the singlecell level: Distribution of secondary metabolites of Arabidopsis thaliana and Hypericum species Core region and lipid A components of lipopolysaccharides Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) application in carbohydrate analysis Glycoproteomic analysis of WGAbound glycoprotein biomarkers in sera from patients with lung adenocarcinoma Synthesis, characterization and in vitro pharmacology of novel pregabalin derivatives An efficient approach to the discovery of potent inhibitors against glycosyltransferases Electrospray and MALDI mass spectrometry: Fundamentals, instrumentation, practicalities, and biological applications Design, synthesis, and immunochemical characterization of a chimeric glycopeptide corresponding to the Shigella flexneri Y Opolysaccharide and its peptide mimic MDWNMHAA Antiproliferative cardenolide glycosides of Elaeodendron alluaudianum from the Madagascar rainforest The influence of microwave irradiation on stereospecific Mo(VI)-catalyzed transformation of deoxysugars A new approach to Amadori ketoses via Mo VIcatalyzed stereospecific isomerization of 2-C-branched sugars bearing azido function in a microwave field Substrate specificity and catalytic mechanism of a xyloglucan xyloglucosyl transferase HvXET6 from barley (Hordeum vulgare L.) Determination of N-glycosylation site and glycan structures of pectin methylesterase in jelly fig (Ficus awkeotsang) achenes The composition of polysaccharides in primary walls of Litchi chinensis Sonn Xyloglucans of monocotyledons have diverse structures The effect of acid dextrinisation on enzyme-resistant starch content in extruded maize starch New oligosaccharides prepared by acid hydrolysis of the polysaccharides from Nerium indicum Mill and their anti-angiogenesis activities Lectin microarray Widely applicable deprotection method of 2,2,2-trichloroethoxycarbonyl (Troc) group using tetrabutylammonium fluoride Hydrolytically stable bioactive synthetic glycopeptide homo-and copolymers by combination of NCA polymerization and click reaction Chemoenzymatic synthesis and lectin array characterization of a class of N-glycan clusters Expeditious chemoenzymatic synthesis of CD52 glycopeptide antigens Arthrobacter endo-b-N-acetylglucosaminidase shows transglycosylation activity on complex-type N-glycan oxazolines: One-pot conversion of ribonuclease B to sialylated ribonuclease C Glycosynthases enable a highly efficient chemoenzymatic synthesis of N-glycoproteins carrying intact natural N-glycans Synthesis of oleanolic acid saponins mimicking components of Chinese folk medicine Di Wu lipID-a software tool for automated assignment of lipids in mass spectra A lipid profile typifies the Beijing strains of Mycobacterium tuberculosis. Identification of a mutation responsible for a modification of the structures of phthiocerol dimycocerosates and phenolic glycolipids IgG glycosylation analysis Supramolecular assembly of carbohydrate-functionalized salphen-metal complexes Synthesis and application of epoxy starch derivatives An Echinococcus multilocularis coproantigen is a surface glycoprotein with unique O-gycosylation Glycoproteomics in neurodegenerative diseases Structural elucidation of a novel B. cenocepacia ET-12 lipooligosaccharide isolated from a cystic fibrosis patient after lung transplantation Against the rules: A marine bacterium, Loktanella rosea, possesses a unique lipopolysaccharide First structural characterization of Burkholderia vietnamiensis lipooligosaccharide from cystic fibrosis-associated lung transplantation strains Fucosylation of chitooligosaccharides by human a1,6-fucosyltransferase requires a nonreducing terminal chitotriose unit as a minimal structure C-Mannosylated peptides derived from the thrombospondin type 1 repeat interact with Hsc70 to modulate its signaling in RAW264.7 cells Two mechanisms of the enhanced antibody-dependent cellular cytotoxicity (ADCC) efficacy of non-fucosylated therapeutic antibodies in human blood Production of a recombinant mouse monoclonal antibody in transgenic silkworm cocoons Homotropic cooperativity of cyclodextrin dimer as an artificial hydrolase Glycomimicry: Display of the GM3 sugar epitope on Escherichia coli and Salmonella enterica sv Typhimurium Enzymatic conversion of diacetylated ANALYSIS OF CARBOHYDRATES AND GLYCOCONJUGATES & sophoroselipid into acetylated glucoselipid: Surface-active properties of novel bolaform biosurfactants Preparation and conformational analysis of C-glycosyl b 2 -and b/b 2 -peptides The multiplicity of N-glycan structures of novine milk 18 kDa lactophorin (Milk GlyCAM-1) Synthesis of an octasubstituted galactose zinc(II) phthalocyanine Anomerically glycosylated zinc(II) naphthalocyanines Synthesis of octaglycosylated Zinc(II) phthalocyanines. Synthesis Sesquiterpene coumarins from Ferula gumosa Transglycosylation-based fluorescent labeling of 6-gala series glycolipids by endogalactosylceramidase Molecular cloning and expression of a novel cholinephosphotransferase involved in glycoglycerophospholipid biosynthesis of Mycoplasma fermentans Enzymatic synthesis of Mycoplasma fermentans specific glycoglycerophospholipid from 1,2-dipalmitoylglycerol Isolation and structural determination of reducing fructooligosaccharides newly produced in stored edible burdock Antiallergic potential of oligomannose-coated liposome-entrapped Cry j 1 as immunotherapy for Japanese cedar pollinosis in mice Enzymatic activity and substrate specifcity of recombinant tomato b-galactosidases 4 and 5 Development of highly efficient and stereocontrolled O-glycosylation methodologies and its application to the construction of bacterial glycans Effects of frozen conditions on stereoselectivity and velocity of O-glycosylation reactions Specificity analysis of lectins and antibodies using remodeled glycoproteins Production, purification and structural characterization of an exopolysaccharide produced by a probiotic Lactobacillus plantarum MTCC 9510 In vitro and in vivo enzymatic syntheses and mass spectrometric database for Nglycans and O-glycans Strategy for glycoproteomics: Identification of glyco-alteration using multiple glycan profiling tools Enrichment strategies for glycopeptides Photooxidative mineralization of microorganisms-produced glycolipid biosurfactants by a titania-mediated advanced oxidation process Highly oriented recombinant glycosyltransferases: Sitespecific immobilization of unstable membrane proteins by using Staphylococcus aureus sortase A A first total synthesis of ganglioside HLG-2 Identification of a glycosylphosphatidylinositol anchormodifying b1-3 N-acetylglucosaminyl transferase in Trypanosoma brucei Direct profiling of tissue lipids by MALDI-TOFMS Glycation sites in neoglycoglycoconjugates from the terminal monosaccharide antigen of the O-PS of Vibrio cholerae O1, serotype Ogawa, and BSA revealed by matrix-assisted laser desorption-ionization tandem mass spectrometry Contribution of Porphyromonas gingivalis lipopolysaccharide to periodontitis Synthesis of new sulfonic acid-containing oligosaccharide mimetics of sialyl Lewis A Synthesis, regioselective hydrogenolysis, partial hydrogenation, and conformational study of dioxane and dioxolanetype (9 0 -anthracenyl)methylene acetals of sugars Mass spectrometric quantification of neutral and sialylated N-glycans from a recombinant therapeutic glycoprotein produced in the two Chinese hamster ovary cell lines Method development for direct detection of glycoproteins on aminophenylboronic acid functionalized self-assembled monolayers by matrix-assisted laser desorption/ionization mass spectrometry Antibody glycans characterization Structural characterization of antibodies by mass spectrometry Mucin-lectin interactions assessed by flow cytometry Mucin-type O-glycosylation-Putting the pieces together Carboxymethylated cyclosophoraose as a novel chiral additive for the stereoisomeric separation of some flavonoids by capillary electrophoresis Characterization of the Streptococcus pneumoniae BgaC protein as a novel surface b-galactosidase with specific hydrolysis activity for the Galb1-3GlcNAc moiety of oligosaccharides Characterization of N-Linked protein glycosylation in Helicobacter pullorum A strategy for precise and large scale identification of core fucosylated glycoproteins Self-assembly of amphiphilic perylenecyclodextrin conjugate and vapor sensing for organic amines Structures of chitosan and chitooligosaccharides and their adsorption toward radionuclide uranium A high-throughput purification of monoclonal antibodies from glycoengineered Pichia pastoris A bifunctional enzyme in a single gene catalyzes the incorporation of GlcN into the Aeromonas core lipopolysaccharide Genetics and proteomics of Aeromonas salmonicida lipopolysaccharide core biosynthesis Selective monitoring of rutin and quercetin based on a novel multi-wall carbon nanotube-coated glassy carbon electrode modified with microbial carbohydrates a-cyclosophorohexadecaose and succinoglycan monomer M3 Utilizing the O-antigen lipopolysaccharide biosynthesis pathway in Escherichia coli to interrogate the substrate specificities of exogenous glycosyltransferase genes in a combinatorial approach Design, synthesis, and evaluation of N-acyl modified sialic acids as inhibitors of adenoviruses causing epidemic keratoconjunctivitis Natural phosphoryl and acyl variants of Lipid A from Neisseria meningitidis strain 89I differentially induce tumor necrosis factor-a in human monocytes Profiles of structural heterogeneity in native lipooligosaccharides of Neisseria and cytokine induction Golgi targeting of Drosophila melanogaster b4GalNAcTB requires a DHHC protein family-related protein as a pilot Strong IgG antibody responses to Borrelia burgdorferi glycolipids in patients with Lyme arthritis, a late manifestation of the infection Generation and characterization of monoclonal antibody to ginsenoside Rg3 A novel, simple and sensitive ligand affinity capture method for detecting molecular interactions by MALDI mass spectrometry GlycoViewer: A tool for visual summary and comparative analysis of the glycome Mass spectrometric molecular-weight determination of highly acidic compounds of biological significance via their complexes with basic polypeptides Enzymatic synthesis of salicin glycosides through transglycosylation catalyzed by amylosucrases from Deinococcus geothermalis and Neisseria polysaccharea Direct analysis of cellulose in poplar stem by matrix-assisted laser desorption/ionization imaging mass spectrometry Superior in vivo efficacy of afucosylated trastuzumab in the treatment of HER2-amplified breast cancer Benzyl ethers as nucleophiles: From hydroxy cyclooctanes toward bridged C-furanosides Multifunctional ZrO 2 nanoparticles and ZrO 2 -SiO 2 nanorods for improved MALDI-MS analysis of cyclodextrins, peptides, and phosphoproteins A synthetic vaccine consisting of a tumor-associated sialyl-TN-MUC1 tandem-repeat glycopeptide and tetanus toxoid: Induction of a strong and highly selective immune response Fully synthetic vaccines consisting of tumor-associated MUC1 glycopeptides and a lipopeptide ligand of the toll-like receptor 2 Ghrelin-like peptide with fatty acid modification and O-glycosylation in the red stingray, Dasyatis akajei Synthesis of glycopeptides Reactivity of rare sugar D-allose during glycation of human serum albumin Arabidopsis thaliana ALG3 mutant synthesizes immature oligosaccharides in the ER and accumulates unique N-glycans Two Arabidopsis thaliana Golgi a-mannosidase I enzymes are responsible for plant N-glycan maturation A sensitive liquid chromatography/mass spectrometry-based assay for quantitation of amino-containing moieties in lipid A Selfpoisoning of Mycobacterium tuberculosis by targeting GlgE in an aglucan pathway Definitive evidence that a single N-glycan among three glycans on inducible costimulator is required for proper protein trafficking and ligand binding AglJ adds the first sugar of the Nlinked pentasaccharide decorating the Haloferax volcanii S-layer glycoprotein ABA-and BAB-triblock cooligomers of tri-O-methylated and unmodified cello-oligosaccharides: Syntheses and structure-solubility relationship Lectin-based enrichment method for glycoproteomics using hollow fiber flow field-flow fractionation: Application to Streptococcus pyogenes Synthesis and characterization of hydroquinone fructoside using Leuconostoc mesenteroides levansucrase High-throughput solid-phase permethylation of glycans prior to mass spectrometry Solid-phase permethylation of glycans for mass spectrometric analysis Antibacterial activity of a disaccharide isolated from Streptomyces sp. strain JJ45 against Xanthomonas sp Insight into the regulation of glycan synthesis in Drosophila chaoptin based on mass spectrometry A tandem mass spectrometric approach to the identification of O-glycosylated glargine glycoforms in active pharmaceutical ingredient expressed in Pichia pastoris Mass spectrometric analysis of carbohydrates labeled with a biotinylated tag Localization of the attachment site of oligoglucans to Mesorhizobium loti HAMBI 1148 murein Synthesis of a heptasaccharide fragment of the mannan from Candida guilliermondii cell wall and its conjugate with BSA Synthesis of 3,6-branched oligomannoside fragments of the mannan from Candida albicans cell wall corresponding to the antigenic factor 4 Reduction of N-linked xylose and fucose by expression of rat b1,4-N-acetylglucosaminyltransferase III in tobacco BY-2 cells depends on Golgi enzyme localization domain and genetic elements used for expression A small-scale method for the preparation of plant N-linked glycans from soluble proteins for analysis by MALDI-TOF mass spectrometry Analyses of biologically active steroids: Antitumor active OSW-1 and cardiotonic marinobufotoxin, by matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight tandem mass spectrometry Structural analogues of diosgenyl saponins: Synthesis and anticancer activity Pyruvoyl, a novel amino protecting group on the solid phase peptide synthesis and the peptide condensation reaction Chemical synthesis of mouse pro-opiomelanocortin(1-74) by azido-protected glycopeptide ligation via the thioester method Correct disulfide pairing is required for the biological activity of crustacean androgenic gland hormone (AGH): Synthetic studies of AGH Loss of UDP-GalNAc: polypeptide N-acetylgalactosaminyltransferase 3 and reduced Oglycosylation in colon carcinoma cells selected for hepatic metastasis Development of tetraphenylethylene-based fluorescent oligosaccharide probes for detection of influenza virus Further structural study of the xyloglucanase-derived eggplant xyloglucan oligo-saccharides Initiation of methylglucose lipopolysaccharide biosynthesis in mycobacteria Synthesis of a chito-tetrasaccharide b-1,4-GlcNAc-b-1,4-GlcN repeating unit A novel polycondensation method for the synthesis of a two-faced b-1,4-glucan Solid-phase synthesis of O-sulfated glycopeptide by the benzylprotected glycan strategy Salmonella enterica serovar Typhimurium lipopolysaccharide deacylation enhances its intracellular growth within macrophages Influence of crystalline forms of titania on desorption/ionization efficiency in titania-based surfaceassisted laser desorption/ionization mass spectrometry Alternative procedures for analysis of lipid A modification with phosphoethanolamine or aminoarabinose Recognition of endogenous ligands by Ctype lectins: Interaction of serum mannan-binding protein with tumorassociated oligosaccharide epitopes Highly fucosylated N-glycan ligands for mannan-binding protein expressed specifically on CD26 (DPPVI) isolated from a human colorectal carcinoma cell line Structural analysis of two trisaccharides isolated from fermented beverage of plant extract Phosphorylase-catalyzed N-formyl-a-glucosaminylation of maltooligosaccharides Gold-catalyzed glycosidations: Unusual cleavage of the interglycosidic bond while studying the armed/ disarmed effect of propargyl glycosides Dendrimers of vaccines consisting of tumor-associated glycopeptide antigens and T cell epitope peptides Bacillus anthracis surfacelayer proteins assemble by binding to the secondary cell wall polysaccharide in a manner that requires csaB and tagO Sialic acids acquired by Pseudomonas aeruginosa are involved in reduced complement deposition and siglec mediated hostcell recognition Mass spectrometric analysis of sulfated N-and Oglycans Steroidal monoglycosides from the Far Eastern starfish Hippasteria kurilensis and hypothetic pathways of polyhydroxysteroid biosynthesis in starfish Two new steroid glycosides from the Far East starfish Hippasteria kurilensis Lectin biosensing using digital analysis of Ru(II)-glycodendrimers Ru(II) glycodendrimers as probes to study lectin-carbohydrate interactions and electrochemically measure monosaccharide and oligosaccharide concentrations Enzymatic synthesis and characterization of hydroquinone galactoside using Kluyveromyces lactis lactase Click synthesis of estradiol-cyclodextrin conjugates as cell compartment selective estrogens Grandidentatin isomer from bark of suwon poplar (Populus alba L.  Populus glandulosa Uyeki) Biotransformation of mulberroside A from Morus alba results in enhancement of tyrosinase inhibition Glycosylation pattern of humanized IgG-like bispecific antibody produced by recombinant CHO cells Mass spectrometric analysis of the glycosphingolipid-derived glycans from miniature pig endothelial cells and islets: Identification of NeuGc epitope in pig islets Rapid and high-throughput analysis of N-glycans from ovarian cancer serum using a 96-well plate platform Identification of a-Gal and non-Gal epitopes in pig corneal endothelial cells and keratocytes by using mass spectrometry Qualitative and quantitative comparison of N-glycans between pig endothelial and islet cells by highperformance liquid chromatography and mass spectrometry-based strategy Construction of a fusion enzyme of dextransucrase and dextranase: Application for onestep synthesis of isomalto-oligosaccharides Enzymatic synthesis of alkyl glucosides using Leuconostoc mesenteroides dextransucrase Alteration in the glycan pattern of pilin in a nonmotile mutant of Synechocystis sp. PCC 6803 Suppression of b-N-acetylglucosaminidase in the N-glycosylation pathway for complex glycoprotein formation in Drosophila S2 cells Carbohydrate moieties contribute significantly to the physicochemical properties of French bean 7S globulin phaseolin Structural characterization of multibranched oligosaccharides from seal milk by a combination of off-line high-performance liquid chromatographymatrix-assisted laser desorption/ionization-time-of-flight mass spectrometry and sequential exoglycosidase digestion Phenylethyl glycosides from Digitalis lanata Acceptor and substrate specificity of b-glucuronidase with transglycosylation activity from Aspergillus niger Kinetic study on the binding of lectin to mannose residues in a polymer brush High molecular weight polyglycerol-based multivalent mannose conjugates 0 -Methyl and 1 0 -xylosyl derivatives of 2 0 -hydroxyflexixanthin are major carotenoids of Hymenobacter species Modifications of human total serum N-glycome during liver fibrosis-cirrhosis, is it all about immunoglobulins? Immunoglobulins are the major glycoproteins involved in the modifications of total serum N-glycome in cirrhotic patients Enrichment of O-GlcNAc modified proteins by the periodate oxidation-hydrazide resin capture approach Surface grafting of cellulose nanocrystals with poly(ethylene oxide) in aqueous media A quantitative model of ultraviolet matrix-assisted laser desorption/ionization A quantitative model of ultraviolet matrix-assisted laser desorption/ionization including analyte ion generation A bipolar rate equation model of MALDI primary and secondary ionization processes, with application to positive/ negative analyte ion ratios and suppression effects Electrospray and MALDI mass spectrometry: Fundamentals, instrumemtation, practialities and biological applications Molecular dynamics simulations of MALDI: Laser fluence and pulse width dependence of plume characteristics and consequences for matrix and analyte ionization Enzymatic polymer synthesis: An opportunity for green polymer chemistry Synthetic studies on the carbohydrate moiety of the antigen from the parasite Echinococcus multilocularis Mass spectrometry for the analysis of milk oligosaccharides Amphiphilic block copolymers based on cyclodextrin host-guest complexes via RAFTpolymerization in aqueous solution Synthesis and conformational analysis of (a-D-galactosyl)phenylmethane and a-,b-difluoromethane analogues: Interactions with the plant lectin viscumin Structures of aldouronic acids liberated from kenaf xylan by endoxylanases from Streptomyces sp Study on systematizing the synthesis of the a-series ganglioside glycans GT1a, GD1a, and GM1 using the newly developed N-Troc-protected GM3 and GalN intermediates Characterization of sialylated and fucosylated glycopeptides of b2-glycoprotein I by a combination of HILIC LC and MALDI MS/MS Glycopeptide profiling of beta-2-glycoprotein I by mass spectrometry reveals attenuated sialylation in patients with antiphospholipid syndrome Application of deep sea yeast-Production of biosurfactants Biosurfactant-producing yeast isolated from Calyptogena soyoae (deep-sea cold-seep clam) in the deep sea An arginyl residue in rice UDP-arabinopyranose mutase is required for catalytic activity and autoglycosylation Structural identification of the main ellagitannins of a boysenberry (Rubus loganbaccus  baileyanus Britt.) extract by LC-ESI-MS/MS, MALDI-TOF-MS and NMR spectroscopy Quantification and characterization of enzymatically produced hyaluronan with fluorophore-assisted carbohydrate electrophoresis Block synthesis of blood group tetrasaccharides B (types 1, 3 and 4) Advances in the separation, sensitive detection, and characterization of heparin and heparan sulfate Glucosinolateaccumulating S-cells in Arabidopsis leaves and flower stalks undergo programmed cell death at early stages of differentiation Polysaccharide microarrays for high-throughput screening of transglycosylase activities in plant extracts Synthesis of a neoglycoconjugate containing a Chlamydophila psittaci-specific branched Kdo trisaccharide epitope The mitA gene of Aspergillus fumigatus is required for mannosylation of inositol-phosphorylceramide, but is dispensable for pathogenicity Sulfated oligosaccharides: New targets for drug development Stability of 5 (6)-carboxyfluorescein in microwave-assisted synthesis of fluoresceinlabelled O-dimannosylated peptides Triterpene glycosides from Agrostemma gracilis Porphyrin-cyclodextrin conjugates as a nanosystem for versatile drug delivery and multimodal cancer therapy A mathematical model of N-linked glycosylation A mathematical model to derive N-glycan structures and cellular enzyme activities from mass spectrometric data Glycosyl azides: Novel substrates for enzymatic transglycosylations Multidimensional system enabling deglycosylation of proteins using a capillary reactor with peptide-Nglycosidase F immobilized on a porous polymer monolith and hydrophilic interaction liquid chromatography-mass spectrometry of glycans Glycomic analysis: An array of technologies Synthesis of an Fmoc-threonine bearing core-2 glycan: A building block for PSGL-1 via Fmoc-assisted solid-phase peptide synthesis The development of retrosynthetic glycan libraries to profile and classify the human serum N-linked glycome Human serum processing and analysis methods for rapid and reproducible N-glycan mass profiling Shigella sonnei oligosaccharide-protein conjugates Immunochemical studies of Shigella flexneri 2a and 6, and Shigella dysenteriae type 1 O-specific polysaccharide-core fragments and their protein conjugates as vaccine candidates a,a-Trehalose-based polyacetals and macrocyclic acetals Application of spectroscopic methods for structural analysis of chitin and chitosan The molecular basis of inhibition of Golgi a-mannosidase II by mannostatin A Glycomics and proteomics analyses of mouse uterine luminal fluid revealed a predominance of Lewis Y and X epitopes on specific protein carriers Productionof sophorolipid biosurfactants by multiple species of the Starmerella (Candida) bombicola yeast clade Four new triterpenes from Anchusa azurea var. azurea Sialyltransferases of marine bacteria efficiently utilize glycosphingolipid substrates Dethreading of deoxyribonucleotides through a-cyclodextrin Efficient guest inclusion by b-cyclodextrin attached to the ends of DNA oligomers upon hybridization to various DNA conjugates Determination of sialic acid and gangliosides in biological samples and dairy products: A review Solid-phase thermodynamic interpretation of ion desorption in matrix-assisted laser desorption/ionization Controlled release of 2,3-desulfated heparin exerts its anti-inflammatory activity by effectively inhibiting E-selectin Deletion of UDP-glucose pyrophosphorylase reveals a UDP-glucose independent UDP-galactose salvage pathway in Leishmania major Imaging of lipids in spinal cord using intermediate pressure matrix-assisted laser desorption-linear ion trap/orbitrap MS Utilization of the linear mode of MALDI-TOF mass spectrometry in the study of glycation during the malting process Profiling of N-linked oligosaccharides using phenylhydrazine derivatization and mass spectrometry Labelling saccharides with phenylhydrazine for electrospray and matrix-assisted laser desorption-ionization mass spectrometry Matrix-assisted laser desorption/ ionization tandem mass spectrometry and post-source decay fragmentation study of phenylhydrazones of N-linked oligosaccharides from ovalbumin Method for investigation of oligosaccharides using phenylhydrazine derivatization N-glycomic changes in human breast carcinoma MCF-7 and T-lymphoblastoid cells after treatment with Herceptin and Herceptin/Lipoplex Mass spectrometric study of N-glycans from serum of woodchucks with liver cancer Chemical characterization, antiproliferative and antiadhesive properties of polysaccharides extracted from Pleurotus pulmonarius mycelium and fruiting bodies Polymerizable vancomycin derivatives for bactericidal biomaterial surface modification: Structure-function evaluation The CHAP domain of Cse functions as an endopeptidase that acts at mature septa to promote Streptococcus thermophilus cell separation Biorecognition of chemically modified bovine serum albumin with lactose prepared under different conditions Conjugates of bovine serum albumin with chitin oligosaccharides prepared through the Maillard reaction Characterization of 4-a-glucanotransferase from Synechocystis sp. PCC 6803 and its application to various corn starches Effects of differential glycosylation of glycodelins on lymphocyte survival Targeted molecular imaging of VEGF receptors overexpressed in ischemic microvasculature using chitosan-DC101 conjugates The poplar GT8E and GT8F glycosyltransferases are functional orthologs of Arabidopsis PARVUS involved in glucuronoxylan biosynthesis The F8H glycosyltransferase is a functional paralog of FRA8 involved in glucuronoxylan biosynthesis in Arabidopsis Down-regulation of PoGT47C expression in poplar results in a reduced glucuronoxylan content and an increased wood digestibility by cellulase The Arabidopsis family GT43 glycosyltransferases form two functionally nonredundant groups essential for the elongation of glucuronoxylan backbone Glycosylated neurotensin analogues exhibit sub-picomolar anticonvulsant potency in a pharmacoresistant model of epilepsy Techniques and tactics used in determining the structure of the trimeric ebolavirus glycoprotein Isolation and tandem mass fragmentations of an anti-inflammatory compound from Aralia elata High-throughput quantitative analysis of plant Nglycan using a DNA sequencer Synthesis of selenium nanowires morphologically directed by Shinorhizobial oligosaccharides Core 3 O-glycan synthase suppresses tumor formation and metastasis of prostate carcinoma PC3 and LNCaP cells through down-regulation of a2b1 integrin complex Core2 O-glycan structure is essential for the cell surface expression of sucrase isomaltase and dipeptidyl peptidase-IV during intestinal cell differentiation Synthesis and cellular uptake properties of guanidine-containing molecular transporters built on the sucrose scaffold Tracing the development of structural elucidation of Nglycans Synthesis of undecaprenyl pyrophosphate-linked glycans as donor substrates for bacterial protein Nglycosylation Deficiency of Dol-P-Man synthase subunit DPM3 bridges the congenital disorders of glycosylation with the dystroglycanopathies N-Glycosylation patterns of hemolymph glycoproteins from Biomphalaria glabrata strains expressing different susceptibility to Schistosoma mansoni infection Multivalent binding of carbohydrates by the human a-defensin, HD51 Structural analysis of sulfated glycans by sequential double-permethylation using methyl iodide and deuteromethyl iodide Sequential enrichment of sulfated glycans by strong anion-exchange chromatography prior to mass spectrometric measurements A new allergen from ragweed (Ambrosia artemisiifolia) with homology to Art v 1 from mugwort An unusual galactofuranose lipopolysaccharide that ensures the intracellular survival of toxin-producing bacteria in their fungal host Human L-selectin preferentially binds synthetic glycosulfopeptides modeled after endoglycan and containing tyrosine sulfate residues and sialyl Lewis x in core 2 O-glycans Efficient synthesis and protein conjugation of b-(1-6)-D-N-acetylglucosamine oligosaccharides from the polysaccharide intercellular adhesin Steroid compounds from two Pacific starfish of the genus Evasterias N-Glycosylation site analysis of human platelet proteins by hydrazide affinity capturing and LC-MS/ MS Phosphoethanolamine substitution of lipid A and resistance of Neisseria gonorrhoeae to cationic antimicrobial peptides and complement-mediated killing by normal human serum Infrared multiphoton dissociation mass spectrometry for structural elucidation of oligosaccharides Collision-induced dissociation tandem mass spectrometry for structural elucidation of glycans Pancreatic cancer serum detection using a lectin/ glyco-antibody array method Structure of pleiotrophin-and hepatocyte growth factor-binding sulfated hexasaccharide determined by biochemical and computational approaches Overexpression and topology of bacterial oligosaccharyltransferase PglB Effects of hapten density on the induced antibody repertoire Advances in the glycosylation of milk protein Light-switchable janus [2] rotaxanes based on a-cyclodextrin derivatives bearing two recognition sites linked with oligo(ethylene glycol) MALDI-TOF-MS analysis of small molecules using modified mesoporous material SBA-15 as assisted matrix Preparation of homogenous oligosaccharide chains from glycosphingolipids Mass spectrometry of fluorocarbon-labeled glycosphingolipids Gold(I)-catalyzed glycosylation with glycosyl ortho-alkynylbenzoates as donors: General scope and application in the synthesis of a cyclic triterpene saponin Immunologic glycosphingolipidomics and NKT cell development in mouse thymus Comparison of sample pre-treatments for laser desorption ionization and secondary ion mass spectrometry imaging of Miscanthus  giganteus Glycan array: A powerful tool for glycomics studies Switching of the core structures of glycosphingolipids from globo-and lacto-to ganglio-series upon human embryonic stem cell differentiation The effect of multivalent binding on the lateral phase separation of adhesive lipids 5-N,4-O-carbonyl-7,8,9-tri-O-chloroacetyl-protected sialyl donor for the stereoselective synthesis of a-(2 ! 9)-tetrasialic acid Enhanced expression of b3-galactosyltransferase 5 activity is sufficient to induce in vivo synthesis of extended type 1 chains on lactosylceramides of selected human colonic carcinoma cell lines A new naphthimidazole derivative for saccharide labeling with enhanced sensitivity in mass spectrometry detection I 2 -Catalyzed oxidative condensation of aldoses with diamines: Synthesis of aldo-naphthimidazoles for carbohydrate analysis In vivo protection provided by a synthetic new alpha-galactosyl ceramide analog against bacterial and viral infections in murine models Fabrication of oriented antibody-conjugated magnetic nanoprobes and their immunoaffinity application Characterization of alginate-like exopolysaccharides isolated from aerobic granular sludge in pilot-plant Synthesis of magnetic nanoparticles with immobilized aminophenylboronic acid for selective capture of glycoproteins The Grignard reaction of cyclodextrin-6-aldehydes revisited: A study of the stereoselectivity upon addition of organometallic reagents to aldehydes and ketones A bivalent glycopeptide to target two putative carbohydrate binding sites on FimH En route to photoaffinity labeling of the bacterial lectin FimH Muropeptide rescue in Bacillus subtilis involves sequential hydrolysis by b-N-acetylglucosaminidase and N-acetylmuramyl-Lalanine amidase Incoherent production reactions of positive and negative ions in matrix-assisted laser desorption/ionization Initial ionization reaction in matrix-assisted laser desorption/ionization Isolation and identification of two novel SDS-resistant secreted chitinases from Aeromonas schubertii Norclerodane diterpenoids from rhizomes of Dioscorea bulbifera An apple oligogalactan prevents against inflammation and carcinogenesis by targeting LPS/TLR4/NF-kB pathway in a mouse model of colitis-associated colon cancer Phosphoryl moieties of lipid A from Neisseria meningitidis and N. gonorrhoeae lipooligosaccharides play an important role in activation of both MyD88-and TRIF-dependent TLR4-MD-2 signaling pathways Efficient synthesis of flaccidoside II, a bioactive component of Chinese folk medicine Di Wu Concise synthesis of two natural triterpenoid saponins, oleanolic acid derivatives isolated from the roots of Pulsatilla chinensis Ionic liquids in sample preparation Albizosides D and E, two new cytotoxic triterpene saponins from Albizia chinensis Cytotoxic oleanane triterpene saponins from Albizia chinensis Electrospray ionization mass spectrometry as a critical tool for revealing new properties of snake venom phospholipase A2 18 F-Labeled galacto and PEGylated RGD dimers for PET imaging of a v b 3 integrin expression Alteration of N-glycome in diethylnitrosamine-induced hepatocellular carcinoma mice: A noninvasive monitoring tool for liver cancer Is permethylation strategy always applicable to protein N-glycosylation study? A case study on the O-acetylation of sialic acid in fish serum glycans Microwave-assisted Kochetkov amination followed by permanent charge derivatization: A facile strategy for glycomics Methylamidation for sialoglycomics by MALDI-MS: A facile derivatization strategy for both a2,3-and a2,6-linked sialic acids Investigation of sample preparation artifacts formed during the enzymatic release of N-linked glycans prior to analysis by capillary electrophoresis Supramolecular assembly of perylene bisimide with b-cyclodextrin grafts as a solid-state fluorescence sensor for vapor detection Elevation of sulfatides in ovarian cancer: An integrated transcriptomic and lipidomic analysis including tissue-imaging mass spectrometry Tandem 18 O stable isotope labeling for quantification of N-glycoproteome Plant-expressed recombinant mountain cedar allergen Jun a 1 is allergenic and has limited pectate lyase activity Modular approach to triazole-linked 1,6-a-D-oligomannosides to the discovery of inhibitors of Mycobacterium tuberculosis cell wall synthetase Glycosylation of b2 subunits regulates GABA A receptor biogenesis and channel gating Lipidomic analysis of porcine olfactory epithelial membranes and cilia A versatile and scalable strategy for glycoprofiling bifidobacterial consumption of human milk oligosaccharides The EgMUR3 xyloglucan galactosyltransferase from Eucalyptus grandis complements the mur3 cell wall phenotype in Arabidopsis thaliana Morphological, toxicological and molecular characterization of a benthic Nodularia isolated from Atlantic estuarine environments Enthalpic studies of xyloglucan-cellulose interactions Titania microparticles and nanoparticles as matrixes for in vitro and in situ analysis of small molecules by MALDI-MS Reactivity-based onepot synthesis of immunosuppressive glycolipids from the Caribbean sponge Plakortis simplex Chemical synthesis of a hyaluronic acid decasaccharide A yeast glycoprotein shows highaffinity binding to the broadly neutralizing human immunodeficiency virus antibody 2G12 and inhibits gp120 interactions with 2G12 and DC-SIGN Fatal outcome due to deficiency of subunit 6 of the conserved oligomeric Golgi complex leading to a new type of congenital disorders of glycosylation Two Kdo-heptose regions identified in Hafnia alvei 32 lipopolysaccharide: The complete core structure and serological screening of different Hafnia O Serotypes Structural analysis of the lipid A isolated from Hafnia alvei 32 and PCM 1192 lipopolysaccharides Functional L-lysine dendritic macromolecules as liver-imaging probes Structural characterization of N-linked oligosaccharides of Defibrase from Agikistrodon acutus by sequential exoglycosidase digestion and MALDI-TOF mass spectrometry Studying a cell division amidase using defined peptidoglycan substrates Data mining the PDB for glyco-related data Facile preparation of fluorescent neoglycoproteins using p-nitrophenyl anthranilate as a heterobifunctional linker Synthesis of the pentasaccharide repeating unit of Escherichia coli O128 antigen A b-D-allopyranoside-grafted Ru(II) complex: Synthesis and acid-base and DNA-binding properties New phosphane based on a b-cyclodextrin, exhibiting a solventtunable conformation, and its catalytic properties Structural analysis of the O-specific polysaccharide isolated from Plesiomonas shigelloides O51 lipopolysaccharide The conformational properties of the Glc 3 Man unit suggest conformational biasing within the chaperone-assisted glycoprotein folding pathway Euscaphinin, a new ellagitannin dimer from Euscaphis japonica (THUNB.) KANITZ Study of lysozyme glycation reaction by mass spectrometry and NMR spectroscopy Fut2-null mice display an altered glycosylation profile and impaired BabAmediated Helicobacter pylori adhesion to gastric mucosa AglP is a S-adenosyl-L-methionine-dependent methyltransferase that participates in the N-glycosylation pathway of Haloferax volcanii Manipulation of electrostatic and saccharide linker interactions in the design of efficient glycopolypeptide-based cholera toxin inhibitors Chemical synthesis and proinflammatory responses of monophosphoryl lipid A adjuvant candidates Glucooligosaccharides production from glucan of Leuconostoc mesenteroides NRRL B-742 by microwave assisted hydrolysis Isorhizochalin: A minor unprecedented bipolar sphingolipid of stereodivergent biogenesis from the Rhizochalina incrustata Development of continuous flow type hydrothermal reactor for hemicellulose fraction recovery from corncob Phenolic compounds from Bursera simaruba Sarg. bark: Phytochemical investigation and quantitative analysis by tandem mass spectrometry Iodine-hexamethyldisilane (HMDS)-mediated anomerization of peracetylated 1,2-trans-linked alkyl and aryl glycosides A simple, mild, and regioselective method for the benzylation of carbohydrate derivatives promoted by silver carbonate Efficient synthesis of a fluorescent tripod detection system for pesticides by microwaveassisted click chemistry Highly efficient deletion of FUT8 in CHO cell lines using zinc-finger nucleases yields cells that produce completely nonfucosylated antibodies Rapid de-O-glycosylation concomitant with peptide labeling using microwave radiation and an alkyl amine base Dissection of host cell signal transduction during Acinetobacter baumannii-triggered inflammatory response Consumption of human milk oligosaccharides by gutrelated microbes Analysis of protein posttranslational modifications using mass spectrometry Interactions of lipopolysaccharide and polymyxin studied by NMR spectroscopy Amino-acetone-bridged cyclodextrins-Artificial alcohol oxidases A systematic approach to protein glycosylation analysis: A path through the maze Flavonoids with prolyl oligopeptidase inhibitory activity isolated from Scutellaria racemosa Pers Glyconanoparticles: Polyvalent tools to study carbohydrate-based interactions Glycosylation protects proteins against free radicals generated from toxic xenobiotics Gold manno-glyconanoparticles: Multivalent systems to block HIV-1 gp120 binding to the lectin DC-SIGN Antithrombin Murcia (K241E) causing antithrombin deficiency: A possible role for altered glycosylation A series of 2-O-trifluoromethylsulfonyl-Dmannopyranosides as precursors for concomitant 18 F-labeling and glycosylation by click chemistry Synthesis and delivery activity of new cationic cholesteryl glucosides Mutational deglycosylation of the Fc portion of immunoglobulin G causes Osulfation of tyrosine adjacently preceding the originally glycosylated site Biochemical and immunological characterization of the plantderived candidate human immunodeficiency virus type 1 mucosal vaccine CTB-MPR 649-684 Supported molecular matrix electrophoresis: A new tool for characterization of glycoproteins Development of an apparatus for rapid release of oligosaccharides at the glycosaminoglycan-protein linkage region in chondroitin sulfate-type proteoglycans Synthesis of sialyllactosamine clusters using carbosilane as core scaffolds by means of chemical and enzymatic approaches Novel synthesis of functional mucin glycopeptides containing both N-and O-glycans Functional neoglycopeptides: Synthesis and characterization of a new class of MUC1 glycoprotein models having core 2-based O-glycan and complex-type N-glycan chains A tethering mechanism for length control in a processive carbohydrate polymerization Structural analysis of arabinoxylans isolated from ball-milled switchgrass biomass Extracellular monoenzyme deglycosylation system of 7-O-linked flavonoid brutinosides and its disaccharide transglycosylation activity from Stilbella fimetaria Biodistribution and excretion of monosaccharidealbumin conjugates measured with in vivo near-infrared fluorescence imaging GlycoForm and glycologue: Two software applications for the rapid construction and display of N-glycans from mammalian sources Site-directed mutagenesis to probe catalysis by a Thermobifida fusca b-1,3-glucanase (Lam81A) The mass-mobility correlation redux: The conformational landscape of anhydrous biomolecules Glycomic analysis by capillary electrophoresis-mass spectrometry Solid-phase permethylation for glycomic analysis High-sensitivity analytical approaches to the analysis of N-glycans Quantitative serum glycomics of esophageal adenocarcinoma and other esophageal disease onsets The effect of glycation on foam and structural properties of b-lactoglobulin Structural characterization of glycans on omega-1, a major Schistosoma mansoni egg glycoprotein that drives Th2 responses Proteomic scale high-sensitivity analyses of GPI membrane anchors Glycotyping of Trypanosoma brucei variant surface glycoprotein MITat1 Studies on the boronation of methyl-b-D-cellobioside-a cellulose model Assessment of heat treatment of dairy products by Synthesis of tetraglucosyl-and tetrapolyamine-tetrabenzoporphyrin conjugates for an application in PDT Synthesis of an Fmoc-Asn-heptasaccharide building block and its application to chemoenzymatic glycopeptide synthesis Fluorescent BODIPYlabelled GM1 gangliosides designed for exploring lipid membrane properties and specific membrane-target interactions Helicobacter pylori binding to new glycans based on N-acetyllactosamine Transcriptomic analysis of Arabidopsis developing stems: A close-up on cell wall genes Glycoforms of human endothelial CD34 that bind L-selectin carry sulfated sialyl Lewis x capped O-and N-glycans Chemical structure analysis of starch and cellulose derivatives Genetic and structural characterization of L11 lipooligosaccharide from Neisseria meningitidis serogroup A strains The N-glycolyl form of mouse sialyl Lewis X is recognized by selectins but not by HECA-452 and FH6 antibodies that were raised against human cells 1-(D-Glucopyranosyl-2 0 -deoxy-2 0 -iminomethyl)-2-hydroxybenzene as chemosensor for aromatic amino acids by switch-on fluorescence Ultrahighly sensitive in situ metabolomic imaging for visualizing spatiotemporal metabolic behaviors Rapid and simple solid-phase esterification of sialic acid residues for quantitative glycomics by mass spectrometry Glycoblotting-assisted O-glycomics: Ammonium carbamate allows for highly efficient O-glycan release from glycoproteins Synthesis and absolute structures of Mycoplasma pneumoniae bglyceroglycolipid antigens Multivalent galacto-trehaloses: design, synthesis, and biological evaluation under the concept of carbohydrate modules Promiscuous activity of ER glucosidase II discovered through donor specificity analysis of UGGT Novel rhamnosyltransferase involved in biosynthesis of serovar 4-specific glycopeptidolipid from Mycobacterium avium complex Systematic synthesis and inhibitory activity of haloacetamidyl oligosaccharide derivatives toward cytoplasmic peptide: N-glycanase Structural and functional characterization of recombinant human serum transferrin secreted from Pichia pastoris The cytotoxic activity of Linum grandiflorum leaves New acylated flavone and cyanogenic glycosides from Linum grandiflorum Synthesis of novel gluco-and galacto-functionalized platinum complexes Design of triazoletethered glycoclusters exhibiting three different spatial arrangements and comparative study of their affinities towards PA-IL and RCA 120 by using a DNA-based glycoarray Capsule polysaccharide conjugate vaccine against diarrheal disease caused by Campylobacter jejuni A-Type crystals from dilute solutions of short amylose chains The N-glycosylation of classical swine fever virus E2 glycoprotein extracellular domain expressed in the milk of goat N-Glycosylation pattern of E2 glycoprotein from classical swine fever virus Glycation pattern of peptides condensed with maltose, lactose and glucose determined by ultraviolet matrix-assisted laser desorption/ionization tandem mass spectrometry Regulated and aberrant glycosylation modulate cardiac electrical signaling Synthesis of hyperbranched b-galceramide-containing dendritic polymers that bind HIV-1 rgp120 In vivo biocompatibility and biodegradability of dextrin-based hydrogels Analysis of glycosylation and other post-translational modifications by mass spectrometry Analysis of N-and Olinked glycans from glycoproteins using MALDI-TOF mass spectrometry Glycosylation pattern of brush borderassociated glycoproteins in enterocyte-like cells: Involvement of complex-type N-glycans in apical trafficking Anthocyanin components and mechanism for color development in blue veronica flowers Biochemical characterization of plasma-derived tissue factor pathway inhibitor: Post-translational modification of free, full-length form with particular reference to the sugar chain Isolation of basidiomycetous yeast Pseudozyma tsukubaensis and production of glycolipid biosurfactant, a diastereomer type of mannosylerythritol lipid-B Production of glycolipid biosurfactants, mannosylerythritol lipids, by a smut fungus, 7 NBRC 32730 Production of a novel glycolipid biosurfactant, mannosylmannitol lipid, by Pseudozyma parantarctica and its interfacial properties Coupling of planar chromatography to mass spectrometry Selective binding of RNase B glycoforms by polydopamine-immobilized concanavalin A Glycosylated Cu/Zn-superoxide dismutase from Kluyveromyces yeast, determined by mass spectrometry Chemoenzymatic synthesis of conjugatable oligosialic acids Analysis of recombinant CD24 glycans by MALDI-TOF-MS reveals prevalence of sialyl-T antigen Separation and identification of oligosaccharides labeled with 3-amino-9-ethylcarbazole using high performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry O-Acetylation of peptidoglycan in gramnegative bacteria. Identification and characterization of peptidoglycan O-acetyltransferase in Neisseria gonorrhoeae Identification of the N-glycosylation sites on recombinant bovine CD38 expressed in Pichia pastoris: Their impact on enzyme stability and catalytic activity Efficient synthesis of 4-amino-4-deoxy-L-arabinose and spacerequipped 4-amino-4-deoxy-L-arabinopyranosides by transglycosylation reactions Molecular weight determination of high molecular mass (glyco)proteins using CGE-on-a-chip Study of inclusion complexes of cycloamylose with surfactants by isothermal titration calorimetry Analysis of N-and O-linked protein glycosylation in children with Prader-Willi syndrome Supramolecular nanocycles comprising b-cyclodextrin-click-ferrocene units: Rings of rings of rings Unique cleavage of 2-acetamido-2-deoxy-Dglucose from the reducing end of biantennary complex type oligosaccharides Efficient and systematic synthesis of a small glycoconjugate library having human complex type oligosaccharides Imaging mass spectrometry: Principle and application Application of thin-layer chromatography/infrared matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometry to structural analysis of bacteria-binding glycosphingolipids selected by affinity detection Chemoenzymatic synthesis of a new class of macrocyclic oligosaccharides Advances on the compositional analysis of glycosphingolipids combining thin-layer chromatography with mass spectrometry Shiga toxins, glycosphingolipid diversity, and endothelial cell injury Heparin/heparan sulfate 6-O-sulfatase from Flavobacterium heparinum. Integrated structural and biochemical investigation of enzyme active site and substrate specificity Heparin/heparan sulfate N-sulfamidase from Flavobacterium heparinum. Structural and biochemical investigation of catalytic nitrogen-sulfur bond cleavage Utilizing ion-pairing hydrophilic interaction chromatography solid phase extraction for efficient glycopeptide enrichment in glycoproteomics Anti-influenza virus activity and structure-activity relationship of aglycoristocetin derivatives with cyclobutenedione carrying hydrophobic chains Structural and functional glycosphingolipidomics by glycoblotting with an aminooxy-functionalized gold nanoparticle Synthesis of urea tethered glycosylated amino acids and glycopeptides mediated by DPPA employing Na-Fmoc-Asp/Glu-5-oxazolidinones Toward fully synthetic homogeneous b-human follicle-stimulating hormone (b-hFSH) with a biantennary N-linked dodecasaccharide. Synthesis of b-hFSH with chitobiose units at the natural linkage sites Experiments on the synthesis of carotenoid glycosides Glycomic analyses of glycoproteins in bile and serum during rat hepatocarcinogenesis Transferrin receptordependent cytotoxicity of artemisinin-transferrin conjugates on prostate cancer cells and induction of apoptosis Synthesis, structural analysis and application of novel acarbose fructoside using levansucrase Extraction and characterization of native heteroxylans from delignified corn stover and aspen A strategy for discovery of cancer glyco-biomarkers in serum using newly developed technologies for glycoproteomics Effect of glycosylation on cis/trans isomerization of prolines in IgA1-hinge peptide Proteomic approaches to study structure, functions and toxicity of legume seeds lectins. Perspectives for the assessment of food quality and safety Engineering host cell lines to reduce terminal sialylation of secreted antibodies Purification and partial characterization of Cu/ Zn superoxide dismutase from Kluyveromyces marxianus yeast Experimental evidence of chemical exchange over the b(1 ! 3) glycosidic linkage and hydrogen bonding involving hydroxy protons in hyaluronan oligosaccharides by NMR spectroscopy Synthesis of amino-bridged oligosaccharide mimetics Production of chitin oligosaccharides with different molecular weights and their antioxidant effect in RAW 264.7 cells Genetic analysis of genes involved in synthesis of modified 4-amino-4,6-dideoxyglucose in flagellin of Pseudomonas syringae pv. tabaci Identification of a naturally-occurring 8-[a-D-glucopyranosyl-(1-6)-b-D-glucopyranosyl] daidzein from cultivated kudzu root Novel rhamnolipid biosurfactants produced by a polycyclic aromatic hydrocarbon-degrading bacterium Pseudomonas aeruginosa strain NY3 Novel O-antigen of Hafnia alvei PCM 1195 lipopolysaccharide with a teichoic acid-like structure Structures of two novel, serologically nonrelated core oligosaccharides of Yokenella regensburgei lipopolysaccharides differing only by a single hexose substitution Syntheses and characterisation of novel cyclodextrin vinyl derivatives from cyclodextrin-nitrophenol-derivatives Use of b-cyclodextrins to control the structure of water-soluble copolymers with hydrophobic parts Facile synthesis of b-cyclodextrin-dextran polymers by "click" chemistry Fullerene sugar balls Gold nanoparticle arrangement on viral particles through carbohydrate recognition: A non-cross-linking approach to optical virus detection Enrichment of glycopeptides for glycan structure and attachment site identification Characterization of the quantitative relationship between signalto-noise (S/N) ratio and sample amount on-target by MALDI-TOF MS: Determination of chondroitin sulfate subsequent to enzymatic digestion Differently complex oligosaccharides can be easily identified by matrix-assisted laser desorption and ionization time-of-flight mass spectrometry directly from a standard thin-layer chromatography plate Identification of O-glycosylated decapeptides within the MUC1 repeat domain as potential MHC class I (A2) binding epitopes Mass spectrometric methods for analysis of oligosaccharides in human milk Preparation and characterization of branched bcyclodextrins having a-L-fucopyranose and a study of their functions Reverse thin layer method for enhanced ion yield of oligosaccharides in matrix-assisted laser desorption/ionization Assessing the cluster glycoside effect during the binding of concanavalin A to mannosylated artificial lipid rafts Systematic screens of a Candida albicans homozygous deletion library decouple morphogenetic switching and pathogenicity Onresin click-glycoconjugation of peptoids Mass spectrometry in the analysis of N-linked and O-linked glycans Glycomics profiling of Chinese hamster ovary cell glycosylation mutants reveals N-glycans of a novel size and complexity Mass spectrometric analysis of mutant mice Characterization of the novel human AGE1hn cell line for production of recombinant proteins Production and characterization of monoclonal antibodies specific to lactotriaosylceramide Repairing faulty genes by aminoglycosides: Development of new derivatives of geneticin (G418) with enhanced suppression of diseases-causing nonsense mutations Development of novel aminoglycoside (NB54) with reduced toxicity and enhanced suppression of disease-causing premature stop mutations Enhanced detection and identification of glycopeptides in negative ion mode mass spectrometry Structural characterization of an oligosaccharide made by Neisseria sicca Electrospray and MALDI mass spectrometry: Fundamentals, instrumentation, practicalities and biological applications Microanalysis of plant cell wall polysaccharides Molecular analysis of carbohydrate-antibody interactions: Case study using a Bacillus anthracis tetrasaccharide Endo-b-N-acetylglucosaminidasecatalyzed polymerization of b-Glcp-(1-4)-GlcpNAc oxazoline: A revisit to enzymatic transglycosylation Syntheses and doxorubicininclusion abilities of b-cyclodextrin derivatives with a hydroquinone a-glycoside residue attached at the primary side Synthesis of sialoglycopolypeptide for potentially blocking influenza virus infection using a rat a2,6-sialyltransferase expressed in BmNPV bacmid-injected silkworm larvae Expression, purification, and analyses of glycosylation and disulfide bonds of Stereum purpureum endopolygalacturonase I in Pichia pastoris Chemically responsive supramolecular assemblies of pyrene-b-cyclodextrin dimer Matrix-assisted laser desorption/ionization mass spectrometric analysis of polysulfated-derived oligosaccharides using pyrenemethylguanidine Structual analysis of neutral and acidic xylooligosaccharides from hardwood kraft pulp, and their utilization by intestinal bacteria in vitro Structural study of the N-glycans of intercellular adhesion molecule-5 (telencephalin) An essential epitope of anti-MUC1 monoclonal antibody KL-6 revealed by focused glycopeptide library Simple and conveniently accessible bi-fluorescence-labeled substrates for amylases Syntheses and biological evaluations of carbosilane dendrimers uniformly functionalized with sialyl a (2 ! 3) lactose moieties as inhibitors for human influenza viruses Characterization of endoplasmic reticulum-localized UDP-Dgalactose: Hydroxyproline O-galactosyltransferase using synthetic peptide substrates in Arabidopsis Structural analysis of three novel trisaccharides isolated from the fermented beverage of plant extracts Novel fructopyranose oligosaccharides isolated from fermented beverage of plant extract Convenient approach to access octa-glycosylated porphyrins via "click chemistry N-Glycosylation engineering of lepidopteran insect cells by the introduction of the b1,4-N-acetylglucosaminyltransferase III gene Efficient substitution reaction from cysteine to the serine residue of glycosylated polypeptide: Repetitive peptide segment ligation strategy and the synthesis of glycosylated tetracontapeptide having acid labile sialyl-TN antigens Structural characterization and dynamics of globotetraosylceramide in vascular endothelial cells under TNF-a stimulation Boronic acid lectin affinity chromatography (BLAC). 3. Temperature dependence of glycoprotein isolation and enrichment Physical mechanisms of bacterial survival revealed by combined grazing-incidence X-ray scattering and Monte Carlo simulation Use of FTIR and mass spectrometry for characterization of glycated caseins Artefacts formed by addition of urea to N-linked glycans released with peptide-N-glycosidase F for analysis by mass spectrometry Okicamelliaside, an extraordinarily potent antidegranulation glucoside isolated from leaves of Camellia japonica Quantification of fructooligosaccharides based on the evaluation of oligomer ratios using an artificial neural network Quantitative glycomics Results of the 2010 glycoprotein research group's (gPRG) study on quantitative glycoprotein analysis Click multivalent heterogeneous neoglycoconjugates-Modular synthesis and evaluation of their binding affinities Globular carbosilane dendrimers with mannose groups at the periphery: Synthesis, characterization and toxicity in dendritic cells Biosynthesis and structure of the Burkholderia cenocepacia K56-2 lipopolysaccharide core oligosaccharide. Truncation of the core oligosaccharide leads to increased binding and sensitivity to polymyxin B Switching of polymerization activity of cinnamoyl-a-cyclodextrin Action of a-Dglucosidase from Aspergillus niger towards dextrin and starch Contiguous Ogalactosylation of 4(R)-hydroxy-l-proline residues forms very stable polyproline II helices Microarrays with varying carbohydrate density reveal distinct subpopulations of serum antibodies Synthesis, in vitro and in vivo studies of Gd-DTPA-XDA-D1-Glc(OH) complex as a new potential MRI contrast agent Structural characterization and hypoglycemic effects of arabinogalactan-protein from the tuberous cortex of the white-skinned sweet potato (Ipomoea batatas L.) Comparison of fluorescent labels for oligosaccharides and introduction of a new postlabeling purification method Characterization of minor N-linked glycans on antibodies using endo H release and MALDI-mass spectrometry Methods in molecular biology, glycomics: Methods and protocols The Caenorhabditis elegans bus-2 mutant reveals a new class of O-glycans affecting bacterial resistance Surfactant-assisted lipopolysaccharide conjugation employing a cyanopyridinium agent and its application to a competitive assay Multifaceted approaches including neoglycolipid oligosaccharide microarrays to ligand discovery for malectin Glycoproteomic profile in wine: A 'sweet' molecular renaissance Mass spectrometry based targeted protein quantification: Methods and applications Convergent synthesis of a common pentasaccharide-repeating unit corresponding to the O-specific polysaccharide of Escherichia coli O4: K3, O4: K6, and O4: K12 Synthesis of penta-and hexasaccharide fragments corresponding to the O-antigen of Escherichia coli O150 Analysis of the human seminal plasma glycome reveals the presence of immunomodulatory carbohydrate functional groups Fast access to robust C-sialoside multimers Synthesis, conjugation, and immunological evaluation of the ANALYSIS OF CARBOHYDRATES AND GLYCOCONJUGATES & serogroup 6 pneumococcal oligosaccharides Human IgG1 expression in silkworm larval hemolymph using BmNPV bacmids and its N-linked glycan structure The synthesis and spectral properties of an encapsulated aminoazobenzene dye O-GlcNAcylation disrupts glyceraldehyde-3-phosphate dehydrogenase homo-tetramer formation and mediates its nuclear translocation Immunochemical studies of trehalose-containing major glycolipid from Tsukamurella pulmonis Structural characterization of the major glycolipids from Arthrobacter globiformis and Arthrobacter scleromae Bioconjugation and detection of lactosamine moiety using a1,3-galactosyltransferase mutants that transfer C2-modified galactose with a chemical handle Muropeptides trigger distinct activation profiles in macrophages and dendritic cells Rapid assembly of gp120 oligosaccharide moieties via one-pot glycosidation-deprotection sequences One-pot catalytic glycosidation/Fmoc removal-An iterable sequence for straightforward assembly of oligosaccharides related to HIV gp120 Analysis of the dispersity in carbohydrate loading of synthetic glycoproteins using MALDI-TOF mass spectrometry Solvent-free synthesis of thioglycosides by ball milling Synthesis of mycobacterial triacylated phosphatidylinositol dimannoside containing an acyl lipid chain at 3-O of inositol O-Glycan inhibitors generate arylglycans, induce apoptosis and lead to growth inhibition in colorectal cancer cell lines Detection of carbohydrates and steroids by cation-enhanced nanostructure-initiator mass spectrometry (NIMS) for biofluid analysis and tissue imaging Protein glycosylation analysis with capillary-based electromigrative separation techniques Glycoprotein analysis using protein microarrays and mass spectrometry Synthesis of 2-deoxy cyclic and linear oligosaccharides by oligomerization of monomers Maintaining product titer while replacing undefined components in a CHO culture system Model a-mannoside conjugates: Immunogenicity and induction of candidacidal activity Hydrodynamic properties of cyclodextrin molecules in dilute solutions Koenigs-Knorr glycosylation with neuraminic acid derivatives Synthesis of the core structure of the lipoteichoic acid of Streptococcus pneumoniae De novo glycan structure search with the CID MS/MS spectra of native N-glycopeptides Identification of a novel human UDP-GalNAc transferase with unique catalytic activity and expression profile Molecular engineering of exocytic vesicle traffic enhances the productivity of Chinese hamster ovary cells The vesicletrafficking protein munc18b increases the secretory capacity of mammalian cells Detection of pathogenic Streptococcus suis bacteria using magnetic glycoparticles Rapid screening of lectins for multivalency effects with a glycodendrimer microarray Imaging MS methodology for more chemical information in less data acquisition time utilizing a hybrid linear ion trap-orbitrap mass spectrometer Atmospheric pressure laser desorption/ionization of plant metabolites and plant tissue using colloidal graphite Click multivalent homogeneous neoglycoconjugates-Synthesis and evaluation of their binding affinities Role of lipid A acylation in Yersinia enterocolitica virulence Glycopeptide-preferring polypeptide GalNAc transferase 10 (ppGalNAc T10), involved in mucin-type O-Glycosylation, has a unique GalNAc-O-Ser/Thr-binding site in its catalytic domain not found in ppGalNAc T1 or T2 Steroidal glycosides from the leaves of Ruscus colchicus: Isolation and structural elucidation based on a preliminary liquid chromatography-electrospray ionization tandem mass spectrometry profiling Off-line coupling of microcolumn separations to desorption mass spectrometry Selective discrimination of cyclodextrin diols using cyclic sulfates 2D-HPLC and MALDI-TOF/TOF analysis of barley proteins glycated during brewing Synthesis of cationic derivatives of Quil A and the preparation of cationic immunestimulating complexes (ISCOMs) Cardenolides from Pergularia tomentosa display cytotoxic activity resulting from their potent inhibition of Na þ /K þ -ATPase Structural determination of the O-chain polysaccharide from the haloalkaliphilic Halomonas alkaliantarctica bacterium strain CRSS Structure of the core region from the lipopolysaccharide of Plesiomonas shigelloides strain 302-73 (serotype O1) Structural characterization of the core region from the lipopolysaccharide of the haloalkaliphilic bacterium Halomonas alkaliantarctica strain CRSS The complete structure of the core of the LPS from Plesiomonas shigelloides 302-73 and the identification of its O-antigen biological repeating unit Enhancement of repair of radiation induced DNA strand breaks in human cells by Ganoderma mushroom polysaccharides Structural investigation of an exopolysaccharide substituted with a lactyl ether group produced by Raoultella terrigena Ez-555-6 isolated in the Chernobyl exclusion zone A protein important for antimicrobial peptide resistance, YdeI/OmdA, is in the periplasm and interacts with OmpD/NmpC Role of E-cadherin N-glycosylation profile in a mammary tumor model Influence of glycosylation on the adsorption of Thermomyces lanuginosus lipase to hydrophobic and hydrophilic surfaces Diazo transfer-click reaction route to new, lipophilic teicoplanin and ristocetin aglycon derivatives with high antibacterial and anti-influenza virus activity: An aggregation and receptor binding study High-energy collision induced dissociation of biomolecules: MALDI-TOF/RTOF mass spectrometry in comparison to tandem sector mass spectrometry Polymer structure of commercial hydrolyzable tannins by matrix-assisted laser desorption/ ionization-time-of-flight mass spectrometry Design, synthesis, and evaluation of novel fluoroquinolone-aminoglycoside hybrid antibiotics Cycloartane-type glycosides from Astragalus amblolepis Triterpenoid saponins from Astragalus wiedemannianus Fischer Mechanisms involved during the ultrasonically induced depolymerization of chitosan: Characterization and control Oligonucleotide carbohydrate-centered galactosyl cluster conjugates synthesized by click and phosphoramidite chemistries Substrate recognition and hydrolysis by a fungal xyloglucan-specific family 12 hydrolase The structural elucidation of glycosaminoglycans Chemical modification and biological evaluation of new semisynthetic derivatives of 28,29-didehydronystatin A1 (S44HP), a genetically engineered antifungal polyene macrolide Context-dependent effects of asparagine glycosylation on Pin WW folding kinetics and thermodynamics MALDI-TOF mass spectrometry of naturally occurring mixtures of monorhamnolipids and dirhamnolipids Functionalized C-glycoside ketohydrazones: Carbohydrate derivatives that retain the ring integrity of the terminal reducing sugar Mass spectrometric-based stable isotopic 2-aminobenzoic acid glycan mapping for rapid glycan screening of biotherapeutics Desorption electrospray ionization mass spectrometry of glycosaminoglycans and their protein noncovalent complex HABA-based ionic liquid matrices for UV-MALDI-MS analysis of heparin and heparan sulfate oligosaccharides Mannose-substituted conjugated polyelectrolyte and oligomer as an intelligent energy transfer pair for label-free visual detection of concanavalin A Development of a CE-MS method to analyze components of the potential biomarker vascular endothelial growth factor 165 Lipopeptides produced by a soil Bacillus megaterium strain Mass spectrometric analysis of the S-layer proteins from Clostridium difficile demonstrates the absence of glycosylation Facile synthesis of mercaptophenylboronic acid-functionalized core-shell structure Fe 3 O 4 @C@Au magnetic microspheres for selective enrichment of glycopeptides and glycoproteins A smart glycol-directed nanodevice from rationally designed macroporous materials Secondary disorders of glycosylation in inborn errors of fructose metabolism Online coupling of thin layer chromatography with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: Synthesis and application of a new material for the identification of carbohydrates by thin layer chromatography/matrix free material enhanced laser desorption/ionization mass spectrometry Acylated triterpenoidal saponins and cytokinins from Gleditsia aquatica Click chemistry approach for the synthesis of water-soluble glycodendrimer with triazole as building unit Synthesis and evaluation of N-acetyl-2-amino-2-deoxy-a-D-galactosyl-1-thio-7-oxaceramide, a new analogue of a-D-galactosyl ceramide Mass spectrometry for pectin structure analysis Multiple site-specific in vitro labeling of single-chain antibody The effect of pH on the production of chitinolytic enzymes of Verticillium fungicola in submerged cultures Glycome-DB. org: A portal for querying across the digital world of carbohydrate sequences Iodine-sodium cyanoborohydride-mediated reductive ring opening of 4,6-O-benzylidene acetals of hexopyranosides Glycosylation patterns of HIV-1 gp120 depend on the type of expressing cells and affect antibody recognition The glycosylation of myeloperoxidase The glycosylation and characterization of the candidate Gc macrophage activating factor Controlled synthesis of linear acyclodextrin oligomers using copper-catalyzed Huisgen 1,3-dipolar cycloaddition Two-step synthesis of galactosylated human serum albumin as a targeted optical imaging agent for peritoneal carcinomatosis Handbook of experimental pharmacology 195, doping in sports Synthesis of multivalent glycoconjugates containing the immunoactive LELTE peptide: Effect of glycosylation on cellular activation and natural killing by human peripheral blood mononuclear cells Golgi function and dysfunction in the first COG4-deficient CDG type II patient Mycobacterium abscessus glycopeptidolipids mask underlying cell wall phosphatidyl-myo-inositol mannosides blocking induction of human macrophage TNF-a by preventing interaction with TLR2 Capsule anchoring in Bacillus anthracis occurs by a transpeptidation reaction that is inhibited by capsidin Synthesis of acceptor substrate analogs for the study of glycosyltransferases involved in the second step of the biosynthesis of O-antigen repeating units Identification of N-linked carbohydrates from severe acute respiratory syndrome (SARS) spike glycoprotein Identification of N-glycans from Ebola virus glycoproteins by matrix-assisted laser desorption/ionisation time-of-flight and negative ion electrospray tandem mass spectrometry Synthesis, characterization, and immunogenicity in mice of Shigella sonnei O-specific oligosaccharide-core-protein conjugates 3-Aminoquinoline acting as matrix and derivatizing agent for MALDI MS analysis of oligosaccharides Cross-linked membranes based on acrylated cyclodextrins and polyethylene glycol dimethacrylates for aromatic/aliphatic separation Mycobacterium marinum lipooligosaccharides are unique caryophyllose-containing cell wall glycolipids that inhibit tumor necrosis factor-a secretion in macrophages Structural analysis of an unusual bioactive N-acylated lipo-oligosaccharide LOS-IV in Mycobacterium marinum Exploiting the cross-metathesis reaction in the synthesis of pseudo-oligosaccharides Isolation and characterization of rhamnolipid-producing bacterial strains from a biodiesel facility Characterisation of haptoglobin in the blood plasma of harbour seals (Phoca vitulina) N-Glycan moieties of the crustacean egg yolk protein and their glycosylation sites Synthesis of Lewis X epitopes on plant N-glycans Glycodynamers: Dynamic polymers bearing oligosaccharides residues -Generation, structure, physicochemical, component exchange, and lectin binding properties Oligosaccharide analysis by graphitized carbon liquid chromatography-mass spectrometry 2-Picoline-borane: A non-toxic reducing agent for oligosaccharide labeling by reductive amination Glycan labeling strategies and their use in identification and quantification Optimized workflow for preparation of APTS-labeled N-glycans allowing high-throughput analysis of human plasma glycomes using 48-channel multiplexed CGE-LIF Decreased levels of bisecting GlcNAc glycoforms of IgG are associated with human longevity Antibacterial activity of N-quaternary chitosan derivatives: Synthesis, characterization and structure activity relationship (SAR) investigations The presence of monoglucosylated N196-glycan is important for the structural stability of storage protein, arylphorin Molecular cloning and characterization of trehalose synthase from Thermotoga maritima DSM3109: Syntheses of trehalose disaccharide analogues and NDP-glucoses MALDI TOF-TOF characterization of a light stabilizer polymer contaminant from polypropylene or polyethylene plastic test tubes Quantification of soyasaponins I and bg in Italian lentil seeds by solid-phase extraction (SPE) and high-performance liquid chromatography-mass spectrometry (HPLC-MS) Nutrition analysis by nanoparticle-assisted laser desorption/ionisation mass spectrometry Isolation and characterization of antibodies against three consecutive Tn-antigen clusters from a phage library displaying human single-chain variable fragments O-GlcNAc modification of the extracellular domain of notch receptors Systematic syntheses of influenza neuraminidase inhibitors: A series of carbosilane dendrimers uniformly functionalized with thioglycoside-type sialic acid moieties Supramolecular assemblies of oligothiophene derivatives bearing b-cyclodextrin 1H-1,2,3-Triazol-1-yl thiodigalactoside derivatives as high affinity galectin-3 inhibitors Synthesis and structural characterization of sialic acid-glutamic acid hybrid foldamers as conformational surrogates of a-2,8-linked polysialic acid The remarkable stability of chimeric, sialic acid derived a/d-peptides in human blood plasma Lack of a-xylosidase activity in arabidopsis alters xyloglucan composition and results in growth defects Intact and top-down characterization of biomolecules and direct analysis using infrared matrix-assisted laser desorption electrospray ionization coupled to FT-ICR mass spectrometry Triumfettosterol Id and triumfettosaponin, a new (fatty acyl)-substituted steroid and a triterpenoid 'dimer' bis(b-D-glucopyranosyl) ester from the leaves of wild Triumfetta cordifolia A. Rich. (Tiliaceae) Fluorescence turn-on sensing of lectins with mannose-substituted tetraphenylethenes based on aggregation-induced emission Synthesis of tri-and pentasaccharide fragments corresponding to the O-antigen of Shigella boydii type 6 Biorecognition of Escherichia coli K88 adhesin for glycated porcine albumin Synthesis of a single-molecule L-rhamnose-containing three-component vaccine and evaluation of antigenicity in the presence of anti-Lrhamnose antibodies Glycosylation of liver acute-phase proteins in pancreatic cancer and chronic pancreatitis Support vector machine prediction of N-and O-glycosylation sites using whole sequence information and subcellular localization Silkworm expression and sugar profiling of human immune cell surface receptor, KIR2DL1 High levels of E4-PHA-reactive oligosaccharides: Potential as marker for cells with characteristics of hepatic progenitor cells Highsensitivity analysis of naturally occurring sugar chains, using a novel fluorescent linker molecule Di-tert-butylsilylenedirected a-selective synthesis of p-nitrophenyl T-antigen analogues Analysis of the human cancer glycome identifies a novel group of tumor-associated N-acetylglucosamine glycan antigens The N-glycome of human embryonic stem cells Inhibition of DC-SIGNmediated HIV infection by a linear trimannoside mimic in a tetravalent presentation Chemo-enzymatic synthesis of poly-N-acetyllactosamine (poly-LacNAc) structures and their characterization for CGL2-galectin-mediated binding of ECM glycoproteins to biomaterial surfaces Aziridine ring opening as regio-and stereoselective access to O-glycosyl amino acids and their transformation into O-glycopeptide mimetics Identification of a polyprenylphosphomannosyl synthase involved in the synthesis of mycobacterial mannosides Reductive amination of the lysine N eamino group leads to a bivalent glyco-amino acid building block suited for SPPS Utilizing Staudinger ligation for the synthesis of glycoamino acid building blocks and other glycomimetics Cysteinebased mannoside glycoclusters: Synthetic routes and antiadhesive properties Development of Dictyostelium discoideum is associated with alteration of fucosylated N-glycan structures TLC/ HPTLC with direct mass spectrometric detection: A review of the progress achieved in the last 5 years O-Glycosylation modulates proprotein convertase activation of angiopoietin-like protein 3. Possible role of polypeptide GalNAc-transferase-2 in regulation of concentrations of plasma lipids Inhibition binding studies of glycodendrimer/lectin interactions using surface plasmon resonance Self-assembling carbohydrate-functionalized oligothiophenes Chemical synthesis using enzymatically generated building units for construction of the human milk pentasaccharides sialyllacto-N-tetraose and sialyllacto-N-neotetraose epimer Hydroxypropyl-substituted b-cyclodextrins: Influence of degree of substitution on the thermodynamics of complexation with tauroconjugated and glycoconjugated bile salts Site-specific analysis of Nlinked oligosaccharides of recombinant lysosomal arylsulfatase A produced in different cell lines Synthesis of a GPI anchor module suitable for protein post-translational modification A combined method for producing homogeneous glycoproteins with eukaryotic N-glycosylation Structural basis of multivalent binding to wheat germ agglutinin Mass spectrometric characterization of the surface-associated 42 kDa lipoprotein JlpA as a glycosylated antigen in strains of Campylobacter jejuni Probing the lactose GM3 carbohydrate-carbohydrate interaction with glycodendrimers Identification of N-glycosylation in prolyl endoprotease from Aspergillus niger and evaluation of the enzyme for its possible application in proteomics Rapid and efficient glycoprotein identification through microwave-assisted enzymatic digestion Assigning Nglycosylation sites of glycoproteins using LC/MSMS in conjunction with endo-M/exoglycosidase mixture Glycan analysis by mass spectrometry Immunoglobulin G glycopeptide profiling by matrixassisted laser desorption ionization Fourier transform ion cyclotron resonance mass spectrometry Glycan bioengineering in immunogen design for tumor T antigen immunotargeting Characterisation of cell wall polysaccharides from okra (Abelmoschus esculentus (L.) Moench) Okra pectin contains an unusual substitution of its rhamnosyl residues with acetyl and alpha-linked galactosyl groups Synthesis and characterization of hydroquinone glucoside using Leuconostoc mesenteroides dextransucrase Lipoprotein lipase and hydrofluoric acid deactivate both bacterial lipoproteins and lipoteichoic acids, but platelet-activating factor-acetylhydrolase degrades only lipoteichoic acids A functional carbohydrate chip platform for analysis of carbohydrate-protein interaction Structural and immunological characterization of the N-glycans from the major yellow jacket allergen Ves v 2: The N-glycan structures are needed for the human antibody recognition Identification of glycosyltransferase 8 family members as xylosyltransferases acting on O-glucosylated notch epidermal growth factor repeats Developments and applications of mass microscopy Characterization of a putative 3-deoxy-D-manno-2-octulosonic acid (Kdo) transferase gene from Arabidopsis thaliana Structural characterisation of the pectic polysaccharide rhamnogalacturonan II using an acidic fingerprinting methodology MALDI linear TOF mass spectrometry of PEGylated (glyco) proteins Synthetic glycosides of ent-caurene series containing substituents with benzyl, phenoxyl, and uracyl fragments SnO 2 @Poly(HEMA-co-St-co-VPBA) core-shell nanoparticles designed for selectively enriching glycopeptides followed by MALDI-MS analysis Mass spectrometry based analysis of protein O-glycosylation Synthesis and characterization of watersoluble conjugated glycopolymer for fluorescent sensing of concanavalin A Laser desorption ionization mass spectrometry by using surface plasmon excitation on gold nanoparticle The N-linked oligosaccharide at FcgRIIIa Asn-45: An inhibitory element for high FcgRIIIa binding affinity to IgG glycoforms lacking core fucosylation Monitoring the hydrolysis and transglycosylation activity of aglucosidase from Aspergillus niger by nuclear magnetic resonance spectroscopy and mass spectrometry N-linked glycan analysis of glycoproteins secreted from rice cell suspension cultures under sugar starvation Characterization of receptor proteins using affinity cross-linking with biotinylated ligands Proton sponge: A novel and versatile MALDI matrix for the analysis of metabolites using mass spectrometry 1,8-Bis(dimethylamino)naphthalene: A novel superbasic matrix for matrix-assisted laser desorption/ionization timeof-flight mass spectrometric analysis of fatty acids Acid-base-driven matrixassisted mass spectrometry for targeted metabolomics Phenolic compounds in the leaves of Populus ussuriensis and their antioxidant activities Phenolic compounds from Populus davidiana wood The structure of the carbohydrate backbone of the lipooligosaccharide from the halophilic bacterium Arcobacter halophilus The structure of the carbohydrate backbone of the lipooligosaccharide from an alkaliphilic Halomonas sp Delineation of the roles of FadD22, FadD26 and FadD29 in the biosynthesis of phthiocerol dimycocerosates and related compounds in Mycobacterium tuberculosis Biodegradable biocompatible xyloglucan films for various applications Transparent xyloglucan-chitosan complex hydrogels for different applications Physico chemical properties of aminated tamarind xyloglucan Fractionation of sugar beet pulp by introducing ion-exchange groups The antimicrobial action of low-molar-mass chitosan, chitosan derivatives and chitooligosaccharides on Bifidobacteria Structural details and composition of Trichomonas vaginalis lipophosphoglycan in relevance to the epithelial immune function Determination of distinctive carbohydrate signatures obtained from the Aeromonas hydrophila (chemotype II) core oligosaccharide pinpointing the presence of the 4-O-phosphorylated 5-O-linked Kdo reducing end group using electrospray ionization quadrupole orthogonal time-offlight mass spectrometry and tandem mass spectrometry Biosynthetic origin of the galactosamine substituent of arabinogalactan in Mycobacterium tuberculosis AftD, a novel essential arabinofuranosyltransferase from mycobacteria NMR structure determination of a segmentally labeled glycoprotein using in vitro glycosylation The role of peptides and proteins in melanoidin formation Optimization of matrix conditions for the control of MALDI in-source decay of permethylated glycans High-spatial resolution matrix-assisted laser desorption ionization imaging analysis of glucosylceramide in spleen sections from a mouse model of Gaucher disease A simple cellulose column procedure for selective enrichment of glycopeptides and characterization by nano LC coupled with electron-transfer and high-energy collisional-dissociation tandem mass spectrometry Structure of compositionally simple lipopolysaccharide from marine Synechococcus Synthesis of water-soluble phthalocyanines bearing four or eight D-galactose units Discrimination of isobaric leucine and isoleucine residues and analysis of post-translational modifications in peptides by MALDI in-source decay mass spectrometry combined with collisional cooling Effect of gas pressure and gas type on the fragmentation of peptide and oligosaccharide ions generated in an elevated pressure UV/IR-MALDI ion source coupled to an orthogonal time-of-flight mass spectrometer Formation and structure elucidation of N-(2,3,4-tri-O-acetyl-b-D-glucopyranosyl)-N 0 -acetylthiourea Disaccharide-modified liposomes and their in vitro intracellular uptake Carbohydrate arrays: Recent developments in fabrication and detection methods with applications Discovery of the first series of small molecule H5N1 entry inhibitors Transglycosylation properties of maltodextrin glucosidase (MalZ) from Escherichia coli and its application for synthesis of a nigerose-containing oligosaccharide Fluorescent glycosylamides produced by microscale derivatization of free glycans for natural glycan microarrays Novel fluorescent glycan microarray strategy reveals ligands for galectins Generation of a natural glycan microarray using 9-fluorenylmethyl chloroformate (FmocCl) as a cleavable fluorescent tag Glycan microarray analysis of Ptype lectins reveals distinct phosphomannose glycan recognition Development and application of mass spectrometric methods for the analysis of progranulin N-glycosylation Francisella tularensis blue-gray phase variation involves structural modifications of lipopolysaccharide Oantigen, core and lipid A and affects intramacrophage survival and vaccine efficacy Glycosylation of gp116 and gp64 envelope proteins of yellow head virus of Penaeus monodon shrimp Structural profiling of individual glycosphingolipids in a single thin-layer chromatogram by multiple sequential immunodetection matched with direct IR-MALDI-o-TOF mass spectrometry Software utilities for the interpretation of mass spectrometric data of glycoconjugates: Application to glycosphingolipids of human serum Ionic liquids in analytical chemistry Isolation of b-mannanase from Cocos nucifera Linn haustorium and its application in the depolymerization of b-(1,4)-linked D-mannans N-Glycan trimming by glucosidase II is essential for Arabidopsis development Immuno-MALDI-TOF MS: New perspectives for clinical applications of mass spectrometry A computational approach for deciphering the organization of glycosaminoglycans Synthesis of galactofuranose-based acceptor substrates for the study of the carbohydrate polymerase GlfT2 Analysis of N-glycans in embryonated chicken egg chorioallantoic and amniotic cells responsible for binding and adaptation of human and avian influenza viruses The mechanism of boron mobility in wheat and canola phloem Removal of the outer Kdo from Helicobacter pylori lipopolysaccharide and its impact on the bacterial surface Galectin-4-regulated delivery of glycoproteins to the brush border membrane of enterocyte-like cells Characterization of an immunodominant cancer-specific O-glycopeptide epitope in murine podoplanin (OTS8) Formation of homooligosaccharides using base-promoted glycosylation of unprotected glycosyl fluorides Glycosyltransferase function in core 2-type protein O-glycosylation Xyloglucan endotransglycosylases (XETs) from germinating nasturtium (Tropaeolum majus) seeds: Isolation and characterization of the major form Botulinum neurotoxin serotype D attacks neurons via two carbohydrate-binding sites in a ganglioside-dependent manner The chain length of lignan macromolecule from flaxseed hulls is determined by the incorporation of coumaric acid glucosides and ferulic acid glucosides Analysis of human plasma lipids and soybean lecithin by means of high-performance thin-layer chromatography and matrix-assisted laser desorption/ionization mass spectrometry Chemoenzymatic synthesis of a glycolipid library and elucidation of the antigenic epitope for construction of a vaccine against Lyme disease Acylated cholesteryl galactosides are specific antigens of Borrelia causing Lyme disease and frequently induce antibodies in late stages of disease Detailed N-glycan analysis of mannose receptor purified from murine spleen indicates tissue specific sialylation Synthesis and acid catalysis of cellulose-derived carbon-based solid acid Experimental studies on the ring opening polymerization of p-dioxanone using an Al(O i Pr) 3 -monosaccharide initiator system Glucuronyltransferase activity of KfiC from Escherichia coli strain K5 requires association of KfiA. KfiC and KfiA are essential enzymes for production of K5 polysaccharide, N-acetylheparosan Current imaging mass spectrometry for metabolite molecules Imaging mass spectrometry for visualization of drug and endogenous metabolite distribution: Toward in situ pharmacometabolomes Imaging mass spectrometry protocols for mass microscopy Structure and kinetic investigation of Streptococcus pyogenes family GH38 a-mannosidase Strategic glycan elution map for the production of human-type N-linked oligosaccharides: The case of hen egg yolk and white Comprehensive analysis of N-linked oligosaccharides from eggs of the family Phasianidae Pectic oligosaccharide analysis by fluorophore-assisted carbohydrate electrophoresis Ionic liquids in analytical chemistry DSA affinity glycoproteome of human liver tissue Isolation of N-linked glycopeptides by hydrazine-functionalized magnetic particles Further insight into the roles of the glycans attached to human blood protein C inhibitor Synthesis of 6-PEtN-a-D-GalpNAc-(1 ! 6)-b-D-Galp-(1 ! 4)-b-D-GlcpNAc-(1 ! 3)-b-D-Galp-(1 ! 4)-b-D-Glcp, a Haemophilus influenzae lipopolysacharide structure, and biotin and protein conjugates thereof Computationally and experimentally derived general rules for fragmentation of various glycosyl bonds in sodium adduct oligosaccharides Molecular cloning of pigeon UDP-galactose: b-D-galactoside a1,4-galactosyltransferase and UDP-galactose: b-Dgalactoside b1,4-galactosyltransferase, two novel enzymes catalyzing the formation of Gal-a1-4Gal-b1-4Gal-b1-4GlcNAc sequence Structural analysis of N-glycans from gull egg white glycoproteins and egg yolk IgG Synthesis of a glycosylphosphatidylinositol anchor bearing unsaturated lipid chains Rapid release of N-linked glycans from glycoproteins by pressure-cycling technology Laser desorption/ionization mass spectrometric analysis of small molecules using fullerene-derivatized silica as energy-absorbing material Synthesis of three regioisomers of the pentasaccharide part of the Skp1 glycoprotein of Dictyostelium discoideum b-Glycosidation of sterically hindered alcohols An Aeromonas caviae genomic island is required for both Oantigen lipopolysaccharide biosynthesis and flagellin glycosylation New triterpene glycoside from Cyclamen adzharicum tubers Mycobacterium leprae phenolglycolipid-1 expressed by engineered M. bovis BCG modulates early interaction with human phagocytes Defects in flagellin glycosylation affect the virulence of Pseudomonas syringae pv. tabaci 6605 Mass spectrometric imaging of ginsenosides localization in Panax ginseng root Thermosensitive hydrogels composed of cyclodextrin pseudorotaxanes. Role of [3]pseudorotaxane in the gel formation Hydrogels composed of organic amphiphiles and a-cyclodextrin: Supramolecular networks of their pseudorotaxanes in aqueous media A plant class V chitinase from a cycad (Cycas revoluta): Biochemical characterization, cDNA isolation, and posttranslational modification Distinct features of matrix-assisted 6 mm infrared laser desorption/ionization mass spectrometry in biomolecular analysis Dissociation profile of protonated fucosyl glycopeptides and quantitation of fucosylation levels of glycoproteins by mass spectrometry Loose-fit polyrotaxane composed of g-cyclodextrin and single poly(ethyelene glycol) chain: Making room in g-CD cavity for additional inclusion complexation Optically pure fullerodendron formed by diastereoselective Dielse-Alder reaction Physiological and glycomic characterization of N-acetylglucosaminyltransferase-IVa and -IVb double deficient mice Synthesis and ring-opening polymerizations of novel S-glycooxazolines Enrichment of Amadori products derived from the nonenzymatic glycation of proteins using microscale boronate affinity chromatography Alterations in receptor-binding properties of swine influenza viruses of the H1 subtype after isolation in embryonated chicken eggs TLC blot (Far-Eastern blot) and its applications Epimeric and amino disaccharide analogs as probes of an a-(1-6)-mannosyltransferase involved in mycobacterial lipoarabinomannan biosynthesis Solution conformation of C-linked antifreeze glycoprotein analogues and modulation of ice recrystallization Application of proteomics in biomarker discovery: A primer for the clinician Synthesis of the glycosaminoglycan-protein linkage tetraosyl peptide moieties of betaglycan, which serve as a hexosamine acceptor for enzymatic glycosyl transfer BacA is indispensable for successful Mesorhizobium-Astragalus symbiosis Synthesis of a sialic acid containing complex-type N-glycan on a solid support A combined 6p-azaelectrocyclization/ Staudinger approach to protein and cell engineering: Noninvasive tumor targeting by N-glycan-engineered lymphocytes Noninvasive imaging of dendrimer-type N-glycan clusters: In vivo dynamics dependence on oligosaccharide structure Synthesis of non-natural xyloglucans by polycondensation of 4,6-dimethoxy-1,3,5-triazin-2-yl oligosaccharide monomers catalyzed by endo-b-1,4-glucanase Novel dialkoxytriazine-type glycosyl donors for cellulase-catalysed lactosylation On-plate-selective enrichment of glycopeptides using boronic acid-modified gold nanoparticles for direct MALDI-QIT-TOF MS analysis Concanavalin A-immobilized magnetic nanoparticles for selective enrichment of glycoproteins and application to glycoproteomics in hepatocelluar carcinoma cell line Biomaterials from sugars: Ring-opening polymerization of a carbohydrate lactone Synthesis of a monophosphoryl derivative of Escherichia coli lipid A and its efficient coupling to a tumor-associated carbohydrate antigen Identification of N-glycan serum markers associated with hepatocellular carcinoma from mass spectrometry data Novel neogala-series glycosphingolipids with terminal mannose and glucose residues from Hirsutella rhossiliensis, an aureobasidin Aresistant ascomycete fungus Functional characterization of tlmH in Streptoalloteichus hindustanus E465-94 ATCC 31158 unveiling new insight into tallysomycin biosynthesis and affording a novel bleomycin analog The effect of temperature on the stability of compounds used as UV-MALDI-MS matrix: 2,5-dihydroxybenzoic acid, 2,4,6-trihydroxyacetophenone, a-cyano-4-hydroxycinnamic acid, 3,5-dimethoxy-4-hydroxycinnamic acid, nor-harmane and harmane Towards an integrated proteomic and glycomic approach to finding cancer biomarkers Chemoenzymatic synthesis of multivalent neoglycoconjugates carrying the helminth glycan antigen LDNF Fragment-based development of triazole-substituted O-galactosyl aldoximes with fragmentinduced affinity and selectivity for galectin-3 TEMPO oxidation of gelatinized potato starch results in acid resistant blocks of glucuronic acid moieties Molecular sieves provoke multiple substitutions in the enzymatic synthesis of fructose oligosaccharide-lauryl esters Synthesis of organic-soluble conjugated polyrotaxanes by polymerization of linked rotaxanes Insulated molecular wire with highly conductive p-conjugated polymer core Reaction of the antitumor antibiotic olivomycin I with aryl diazonium salts. Synthesis, cytotoxic and antiretroviral potency of 5-aryldiazenyl-6-Odeglycosyl derivatives of olivomycin I Highly site-selective stability increases by glycosylation of dihydrofolate reductase Gold-catalyzed glycosidations: Synthesis of 1,6-anhydro saccharides Site-specific glycoprofiling of N-linked glycopeptides using MALDI-TOF MS: Strong correlation between signal strength and glycoform quantities Purification and characterization of a new recombinant factor VIII (N8) Identification and quantification of N-linked oligosaccharides released from glycoproteins: An inter-laboratory study Determination of Nlinked sialyl-sugar chains in the lungs of domestic cats and dogs in Thailand susceptible to the highly pathogenic avian influenza virus (H5N1) Cyclodextrin-based hyperbranched polymers: Molecule design, synthesis and characterization Glycoproteomics: Past, present and future Molecular basis for galactosylation of core fucose residues in invertebrates. Identification of Caenorhabditis elegans N-glycan core a1,6-fucoside b1,4-galactosyltransferase galt-1 as a member of a novel glycosyltransferase family Glycoproteomic analysis of abnormal N-glycosylation on the kappa chain of cryocrystalglobulin in a patient of multiple myeloma Solvolytic depolymerization of chondroitin and dermatan sulfates A chromatographic approach for elevating the antibody-dependent cellular cytotoxicity of antibody composites Structural characterization and surface-active properties of a succinoyl trehalose lipid produced by Rhodococcus sp. SD-74 Chemo-enzymatic synthesis of glycosylated insulin using a GlcNAc tag Acceptor specificity in the transglycosylation reaction using Endo-M Coxiella burnetii glycomics and proteomics-Tools for linking structure to function Social self-sorting: Alternating supramolecular oligomer consisting of isomers Humanization of recombinant glycoproteins expressed in insect cells Inactivation of Mycobacterium tuberculosis mannosyltransferase pimB reduces the cell wall lipoarabinomannan and lipomannan content and increases the rate of bacterial-induced human macrophage cell death The recognition motif of the glycoprotein-folding sensor enzyme UDP-Glc: Glycoprotein glucosyltransferase Enrichment method of sulfated glycopeptides by a sulfate emerging and ion exchange chromatography Enzymatic solubilization of brewers' spent grain by combined action of carbohydrases and peptidases Poly(N-vinylpyrrolidone) bearing covalently attached cyclodextrin via click-chemistry: Synthesis, characterization, and complexation behavior with phenolphthalein Chemical and enzymatic N-glycan release comparison for N-glycan profiling of monoclonal antibodies expressed in plants Analysis of carbohydrates on proteins by offline normal-phase liquid chromatography MALDI-TOF/TOF-MS/ MS Carbohydrate structural analysis of wheat flour arabinogalactan protein The fine structure of Neisseria meningitidis lipooligosaccharide from the M986 strain and three of its variants Dihydrobenzoic acid modified nanoparticle as a MALDI-TOF MS matrix for soft ionization and structure determination of small molecules with diverse structures Photoinduced isomerization of allyl alcohols to carbonyl compounds using dendrimer disulfide as catalyst Miniaturizing sample spots for matrix-assisted laser desorption/ionization mass spectrometry Glycomics profiling of heparan sulfate structure and activity Alkali-hydroxide-doped matrices for structural characterization of neutral underivatized oligosaccharides by MALDI time-of-flight mass spectrometry Phosphocholinecontaining glycosyl inositol-phosphoceramides from Trichoderma viride induce defense responses in cultured rice cells Targeted serum glycoproteomics for the discovery of lung cancer-associated glycosylation disorders using lectin-coupled Pro-teinChip arrays Chemoenzymatic synthesis of glycosylated glucagon-like peptide 1: Effect of glycosylation on proteolytic resistance and in vivo blood glucoselowering activity Daughter cell separation is controlled by cytokinetic ring-activated cell wall hydrolysis Solid-phase synthesis of glycopeptide carrying a tetra-N-acetyllactosamine-containing core 2 decasaccharide Glycosylation specific for adhesion molecules in epidermis and its receptor revealed by glycoform-focused reverse genomics Isolation and identification of arabinose mycolates of cell wall skeleton (CWS) derived from Mycobacterium bovis BCG Tokyo 172 (SMP-105) Synthesis, cell-surface binding, and cellular uptake of fluorescently labeled glucose-DNA conjugates with different carbohydrate presentation Use of a novel ionic liquid matrix for MALDI-MS analysis of glycopeptides and glycans out of total tryptic digests Efficient transfer of sialo-oligosaccharide onto proteins by combined use of a glycosynthase-like mutant of Mucor hiemalis endoglycosidase and synthetic sialo-complex-type sugar oxazoline Milk oligosaccharides Structural and functional characterization of a putative polysaccharide deacetylase of the human parasite Encephalitozoon cuniculi Glycosyl conjugates of biotinylated diaminopyridine applied for study of carbohydrate-to-carbohydrate interaction An oxidative enzyme boosting the enzymatic conversion of recalcitrant polysaccharides Deciphering carbohydrate structures by IMS-MS. Applications to biological features related to carbohydrate chemistry and biology Software platform for high-throughput glycomics Preparation and properties of polyurethanes based on castor oil chemically modified with yucca starch glycoside Synthesis and characterization of neurostatin-related compounds with high inhibitory activity of glioma growth O-Fucosylation of an antibody light chain: Characterization of a modification occurring on an IgG1 molecule Asparagine-linked oligosaccharides present on a non-consensus amino acid sequence in the C H 1 domain of human antibodies Biochemical characterization of bovine plasma thrombin-activatable fibrinolysis inhibitor (TAFI) Glycosylation pattern of mature dimeric leukocyte and recombinant monomeric myeloperoxidase. Glycosylation is required for optimal enzymatic activity N-Linked glycosylation is an important parameter for optimal selection of cell lines producing biopharmaceutical human IgG Immunoglobulin G galactosylation and sialylation are associated with pregnancyinduced improvement of rheumatoid arthritis and the postpartum flare: Results from a large prospective cohort study Structural and functional analysis of glycosphingolipids of Schistosoma mansoni Elucidation of molecular diversity and body distribution of saponins in the sea cucumber Holothuria forskali (Echinodermata) by mass spectrometry Qualitative and quantitative saponin contents in five sea cucumbers from the Indian Ocean Localization of secondary metabolites in marine invertebrates: Contribution of MALDI MSI for the study of saponins in cuvierian tubules of H. forskali Elevated sulfatide levels in neurons cause lethal audiogenic seizures in mice Rhizobium leguminosarum biovar viciae 3841, deficient in 27-hydroxyoctacosanoate-modified lipopolysaccharide, is impaired in desiccation tolerance, biofilm formation and motility Glycome profiling using modern glycomics technology: Technical aspects and applications Symbol nomenclature for glycan representation Synthesis of cyclic b-glucan using Laminarinase 16A glycosynthase mutant from the basidiomycete Phanerochaete chrysosporium Purification and Nglycosylation analysis of melanoma antigen dopachrome tautomerase Rapana venosa hemocyanin with antiviral activity Stable isotope-enhanced two-and threedimensional diffusion ordered 13 C NMR spectroscopy (SIE-DOSY 13 C NMR) Modern MALDI time-of-flight mass spectrometry Orthogonal activation of propargyl and n-pentenyl glycosides and 1,2-orthoesters Laser desorption-ionization of lipid transfers: Tissue mass spectrometry imaging without MALDI matrix Characterization of branched polysaccharides using multiple-detection size separation techniques Multigram synthesis of a hyaluronic acid subunit and synthesis of fully protected oligomers Finding new posttranslational modifications in salivary proline-rich proteins Enzymatic glycosylations on arrays Rapid assembly of oligosaccharides: A highly convergent strategy for the assembly of a glycosylated amino acid derived from PSGL-1 Databases and informatics for glycobiology and glycomics Production of nonfucosylated antibodies by co-expression of heterologous GDP-6-deoxy-D-lyxo-4-hexulose reductase Characterization of the acidic N-linked glycans of the zona pellucida of prepuberal pigs by a mass spectrometric approach Pectin, a versatile polysaccharide present in plant cell walls Expression of Talaromyces emersonii cellobiohydrolase Cel7A in Saccharomyces cerevisiae and rational mutagenesis to improve its thermostability and activity Structural characterization of flavonoid glycosides by multi-stage mass spectrometry The absence of an identifiable single catalytic base residue in Thermobifida fusca exocellulase Cel6B Quantitation of saccharide compositions of O-glycans by mass spectrometry of glycopeptides and its application to rheumatoid arthritis Comparison of methods for profiling O-glycosylation. Human Proteome Organisation Human Disease Glycomics/Proteome Initiative multi-institutional study of IgA1 Fluorinated glycosyl amino acids for mucin-like glycopeptide antigen analogues A nonprotein thermal hysteresis-producing xylomannan antifreeze in the freeze-tolerant Alaskan beetle Upis ceramboides Glycans on influenza hemagglutinin affect receptor binding and immune response Synthesis of a mannose-capped disaccharide with a thiol terminus Steroidal saponins from the rhizomes of Polygonatum odoratum Oxidative stress-induced peptidoglycan deacetylase in Helicobacter pylori Convenient synthesis of an N-glycan octasaccharide of the bisecting type Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Application in carbohydrate analysis Molecular aggregation behavior of perylene-bridged bis(b-cyclodextrin) and its electronic interactions upon selective binding with aromatic guests Functional characterization of tlmK unveiling unstable carbinolamide intermediates in the tallysomycin biosynthetic pathway Silver-coated gold nanoparticles as concentrating probes and matrices for surface-assisted laser desorption/ionization mass spectrometric analysis of aminoglycosides Conversion of squid pen by Pseudomonas aeruginosa K187 fermentation for the production of Nacetyl chitooligosaccharides and biofertilizers N-Terminal deletion of peptide: N-Glycanase results in enhanced deglycosylation activity A new chalcone glycoside, a new tetrahydrofuranoid lignan, and antioxidative constituents from the stems and leaves of Viburnum propinquum Cosmc is an essential chaperone for correct protein O-glycosylation Role of a cytoplasmic dual-function glycosyltransferase in O 2 regulation of development in Dictyostelium Crystal structure and statistical coupling analysis of highly glycosylated peroxidase from royal palm tree (Roystonea regia) Two-enzyme systems for glycolipid and polyglycerolphosphate lipoteichoic acid synthesis in Listeria monocytogenes N-Glycan analysis of recombinant L-selectin reveals sulfated GalNAc and GalNAc-GalNAc motifs Glycosylation of the phosphate binding protein, PstS, in Streptomyces coelicolor by a pathway that resembles protein O-mannosylation in eukaryotes Immobilization of enzyme on detonation nanodiamond for highly efficient proteolysis Comparative glycoproteomics: Approaches and applications Covalent modification of chitin with silk-derivatives acts as an amphiphilic self-organizing template in nacre biomineralisation Expression of Helix pomatia lectin binding glycoproteins in women with breast cancer in relationship to their blood group phenotypes Analysis of site-specific glycosylation of renal and hepatic g-glutamyl transpeptidase from normal human tissue Release and characterization of single side chains of white cabbage pectin and their complement-fixing activity Characterization of WbpB, WbpE, and WbpD and reconstitution of a pathway for the biosynthesis of UDP-2,3-diacetamido-2,3-dideoxy-D-mannuronic acid in Pseudomonas aeruginosa MALDI-TOF MS and CE-LIF fingerprinting of plant cell wall polysaccharide digests as a screening tool for Arabidopsis cell wall mutants Branched arabino-oligosaccharides isolated from sugar beet arabinan Derivatization of sialic acids for stabilization in matrix-assisted laser desorption/ionization mass spectrometry and concomitant differentiation of a(2-3) and a(2-6) isomers Glycomic characterization of prostate-specific antigen and prostatic acid phosphatase in prostate cancer and benign disease seminal plasma fluids Synthesis of bifunctional peptide derivatives based on a b-cyclodextrin core with drug delivery potential Glycoprotein production for structure analysis with stable, glycosylation mutant CHO cell lines established by fluorescence-activated cell sorting Triple recognition of B-DNA by a neomycin-Hoechst 33258-pyrene conjugate Introductory glycosylation analysis using SDS-PAGE and peptide mass fingerprinting Synthesis and cytotoxic activity of G3PAMAM-NH 2 dendrimer-modified digoxin and proscillaridin A conjugates in breast cancer cells Quantitative site-specific analysis of protein glycosylation by LC-MS using different glycopeptide-enrichment strategies Profiling of N-glycosylation gene expression in CHO cell fed-batch cultures An investigation of intracellular glycosylation activities in CHO cells: Effects of nucleotide sugar precursor feeding vitro bacterial polysaccharide biosynthesis: Defining the functions of Wzy and Wzz Ultrasound ionization of biomolecules New development of glycan arrays Gold nanoparticles as assisted matrices for the detection of biomolecules in a high-salt solution through laser desorption/ionization mass spectrometry Development of an annotated library of neutral human milk oligosaccharides Synthesis of monomeric and dimeric repeating units of the zwitterionic type 1 capsular polysaccharide from Streptococcus pneumoniae Chemoenzymatic synthesis of glycosylphosphatidylinositol-anchored glycopeptides Synthesis of mangiferin, isomangiferin, and homomangiferin Sortase A-catalyzed transpeptidation of glycosylphosphatidylinositol derivatives for chemoenzymatic synthesis of GPI-anchored proteins Synthesis of talosin A and B, two bioactive isoflavonoid glycosides Direct labelling of peptides with 2-[ 18 F]fluoro-2-deoxy-D-glucose ([ 18 F]FDG) Structural glycomics using hydrophilic interaction chromatography (HILIC) with mass spectrometry Hexose rearrangements upon fragmentation of N-glycopeptides and reductively aminated Nglycans Ligand identification of carbohydrate-binding proteins employing a biotinylated glycan binding assay and tandem mass spectrometry Glycan reductive isotope labeling for quantitative glycomics Core 3-derived O-glycans are essential for intestinal mucus barrier function Rapid comparison of a candidate biosimilar to an innovator monoclonal antibody with advanced liquid chromatography and mass spectrometry technologies Preparation of chitooligosaccharides by the enzymatic hydrolysis of chitosan Potent angiogenic inhibition effects of deacetylated chitohexaose separated from chitooligosaccharides and its mechanism of action in vitro Synthesis of well-defined 7-arm and 21-arm poly(Nisopropylacrylamide) star polymers with b-cyclodextrin cores via click chemistry and their thermal phase transition behavior in aqueous solution Chitooligosaccharides protect human embryonic hepatocytes against oxidative stress nduced by hydrogen peroxide On-plate enrichment of glycopeptides by using boronic acid functionalized gold-coated Si wafer Highly specific enrichment of glycopeptides using boronic acid-functionalized mesoporous silica N-Glycosylation profiling of turtle egg yolk: Expression of galabiose structure Development and application of high performance liquid chromatography map of glucuronyl N-glycans Comparative studies on the structural features of O-glycans between leukemia and epithelial cell lines Hyphenated technique for releasing and MALDI MS analysis of Oglycans in mucin-type glycoprotein samples Preparation and characterization of phospha sugar analogues, 2,3-dibromo-3-methyl-1-phenylphospholane 1-oxide derivatives, as novel anticancer agents Synthesis of b-D-fructopyranosyl-(2 ! 6)-D-glucopyranose from D-glucose and D-fructose by a thermal treatment Switching from altro-a-cyclodextrin dimer to pseudo[1]rotaxane dimer through tumbling A molecular reel: Shuttling of a rotor by tumbling of a macrocycle Selective enrichment of glycopeptides/phosphopeptides using porous titania microspheres Synthesis of di-and tetrasaccharide containing 6-deoxytalose from the O-antigenic polysaccharide of B. pseudomallei strain 304b Mechanism of mild acid hydrolysis of galactan polysaccharides with highly ordered disaccharide repeats leading to a complete series of exclusively oddnumbered oligosaccharides Heterogeneous components of chitosans Glucopyranoside-incorporated N-heterocyclic carbene complexes of silver(I) and palladium(II): Efficient water-soluble Suzuki-Miyaura coupling palladium(II) catalysts Detection of small neutral carbohydrates using various supporting materials in laser desorption/onization mass spectrometry Synthesis of a novel class of glycocluster with a cyclic a-(1 ! 6)-octaglucoside as a scaffold and their binding abilities to concanavalin A Isolation and structural characterization of a polysaccharide FCAP1 from the fruit of Cornus officinalis Expression, glycoform characterization, and antibody-binding of HIV-1 V3 glycopeptide domain fused with human IgG1-Fc Synthesis of kaempferol 3-O-(3 00 ,6 00 -Di-O-E-p-coumaroyl)-b-D-glucopyranoside, efficient glycosylation of flavonol 3-OH with glycosyl odlkynylbenzoates as donors Total synthesis and structural revision of TMGchitotriomycin, a specific inhibitor of insect and fungal b-N-acetylglucosaminidases Chemoselective glycosylation of carboxylic acid with glycosyl ortho-hexynylbenzoates as donors Homogeneous synthesis of GRGDY grafted chitosan on hydroxyl groups by photochemical reaction for improved cell adhesion Facile synthesis of 4-mercaptophenylboronic acid functionalized gold nanoparticles for selective enrichment of glycopeptides New triterpenoid saponins from the roots of Gypsophila paniculata L The crystal structure of a xyloglucan-specific endo-b-1,4-glucanase from Geotrichum sp. M128 xyloglucanase reveals a key amino acid residue for substrate specificity Imidazolium cation supported solution-phase assembly of homolinear a(1 ! 6)-linked octamannoside: An efficient alternate approach for oligosaccharide synthesis Growth phasedependent expression of proteins with decreased plant-specific Nglycans and immunogenicity in tobacco BY2 cells Oxidation of the primary hydroxyl group of galactose of galactaosyl ceramide analogue by chemical method-Precursors for the synthesis of labeled conjugates Developmental changes in glycolipids and synchronized expression of nutrient transporters in the mouse small intestine Differences in N-glycan structures found on recombinant IgA1 and IgA2 produced in murine myeloma and CHO cell lines Extraction of glycosaminoglycan from sea hare, Aplysia kurodai, and its functional properties 2. Structural properties of purified glycosaminoglycan Synthesis of dopamine and l-DOPAa-glycosides by reaction with cyclomaltohexaose catalyzed by cyclomaltodextrin glucanyltransferase N-Glycosylation status of b-haptoglobin in sera of patients with prostate cancer vs. benign prostate diseases Unexpected tolerance of glycosylation by UDP-GalNAc:polypeptide a-N-acetylgalactosaminyltransferase revealed by electron capture dissociation mass spectrometry: Carbohydrate as potential protective groups Modification-specific proteomics in plant biology Effect and limitation of excess ammonium on the release of O-glycans in reducing forms from glycoproteins under mild alkaline conditions for glycomic and functional analysis Hydrophilic interaction chromatography based enrichment of glycopeptides by using click maltose: A matrix with high selectivity and glycosylation heterogeneity coverage Construction and transmembrane dissociation behavior of supramolecular assembly of quinolinocyclodextrin with porphyrin Metabolic labeling of glycoconjugates with photocrosslinking sugars Identification of blood group A/ A-Leb/y and B/B-Leb/y active glycotopes co-expressed on the Oglycans isolated from two distinct human ovarian cyst fluids A supramolecular bifunctional artificial enzyme with superoxide dismutase and glutathione peroxidase activities Enabling techniques and strategic workflow for sulfoglycomics based on mass spectrometry mapping and sequencing of permethylated sulfated glycans Synthesis and 13 C NMR spectroscopy of model compounds for the microstructure analysis of poly(vinyl glycoside)s Production of anti-carbohydrate antibodies by phage display technologies. Potential impairment of cell growth as a result of endogenous expression Precise structure of acidic polysaccharide present in Salvia hydrogels N-glycosylation in Archaea: On the coordinated actions of Haloferax volcanii AglF and AglM On-line separations combined with MS for analysis of glycosaminoglycans Mass spectrometry and glycomics Matrix-assisted laser desorption/ionization imaging mass spectrometry Effect of enzyme processivity on the efficacy of a competitive chitinase inhibitor Catalytic properties of endo-1,3-b-D-glucanase from the Vietnamese edible mussel Perna viridis Cerebrocostomandibular-like syndrome and a mutation in the conserved oligomeric Golgi complex, subunit 1 A new mutation in COG7 extends the spectrum of COG subunit deficiencies Clickable lipids: Azido and alkynyl fatty acids and triacylglycerols Expression, purification, and characterization of recombinant human transferrin from rice Structural characterization and anti-fatigue activity of polysaccharides from the roots of Morinda officinalis A modified synthesis and serological evaluation of neoglycoproteins containing the natural disaccharide of PGL-I from Mycobacterium leprae Synthesis of novel aminoglycosides via allylic azide rearrangement for investigating the significance of 2 0 -amino group Development of a plate-based scintillation proximity assay for the mycobacterial AftB enzyme involved in cell wall arabinan biosynthesis Transient expression and purification of chimeric heavy chain antibodies Extracting both peptide sequence and glycan structural information by 157 nm photodissociation of N-linked glycopeptides Synthesis of Neu5Ac-Gal-functionalized gold glyconanoparticles Total synthesis of apigenin-4 0 -yl-2-O-(pcoumaroyl)-b-D-glucopyranoside Recent developments of nanoparticle-based enrichment methods for mass spectrometric analysis in proteomics Specific enrichment methods for glycoproteome research Boronic acid functionalized core-satellite composite nanoparticles for advanced enrichment of glycopeptides and glycoproteins Condensed tannins from mangrove species Kandelia candel and Rhizophora mangle and their antioxidant activity LEW3, encoding a putative a-1,2-mannosyltransferase (ALG11) in N-linked glycoprotein, plays vital roles in cell-wall biosynthesis and the abiotic stress response in Arabidopsis thaliana A perspective on the Maillard reaction and the analysis of protein glycation by mass spectrometry: Probing the pathogenesis of chronic disease A novel strategy for MALDI-TOF MS analysis of small molecules Steroidal saponins from the rhizomes of Paris delavayi Acute kidney injury induced by proteinoverload nephropathy down-regulates gene expression of hepatic cerebroside sulfotransferase in mice, resulting in reduction of liver and serum sulfatides Prediction of collision-induced dissociation spectra of common N-glycopeptides for glycoform identification Mass spectrometry for structural characterization of therapeutic antibodies Analysis of protein glycosylation and phosphorylation using liquid phase separation, protein microarray technology, and mass spectrometry Preparation and characterization of galacto-mannan-oligosaccharides hydrolyzed from guar gum by bmannanase Inclusion behavior of bcyclodextrin with bipyridine molecules: Factors governing host-guest inclusion geometries Combination of b-elimination and liquid chromatography/quadrupole time-of-flight mass spectrometry for the determination of O-glycosylation sites Study of matrix additives for sensitive analysis of lipid A by matrix-assisted laser desorption ionization mass spectrometry Microwaveassisted sample preparation for rapid and sensitive analysis of H. pylori lipid A applicable to a single colony BC10, a DUF266-containing and Golgi-located type II membrane protein, is required for cell-wall biosynthesis in rice Synthesis of ionic liquids functionalized b-cyclodextrin-bonded chiral stationary phases and their applications in high-performance liquid chromatography Rapid identification of gallotannins from Chinese galls by matrix-assisted laser desorption/ ionization time-of-flight quadrupole ion trap mass spectrometry Engineered nanoparticle surfaces for improved mass spectrometric analysis Convenient synthesis of sulfated oligofucosides Synthesis of a mannose hexasaccharide related to the cell wall mannan of Candida dubliniensis and Trychophyton mentagrophytes Synthesis of the 6-deoxytalose-containing tri-and hexasaccharides of the O-antigen polysaccharide from Mesorhizobium huakuii IFO15243T Highly efficient removal of allyloxycarbonyl (Alloc) function provides a practical orthogonal protective strategy for carbohydrates Dual reversible self-assembly of PNIPAM-based amphiphiles formed by inclusion complexation Cytotoxic triterpenoid saponins acetylated with monoterpenoid acid from Albizia julibrissin