key: cord-0937377-de53rg1o
authors: Su, Wen-Yu; Du, Pin-Xian; Santos, Harvey M.; Ho, Tzong-Shiann; Keskin, Batuhan Birol; Pau, Chi Ho; Yang, An-Ming; Chou, Yi-Yu; Shih, Hsi-Chang; Syu, Guan-Da
title: Antibody Profiling in COVID-19 Patients with Different Severities by Using Spike Variant Protein Microarrays
date: 2022-04-20
journal: Anal Chem
DOI: 10.1021/acs.analchem.1c05567
sha: 89b4cb7b6f178ae6ab286df6f9ee7185e581e2c8
doc_id: 937377
cord_uid: de53rg1o

[Image: see text] The disease progression of COVID-19 varies from mild to severe, even death. However, the link between COVID-19 severities and humoral immune specificities is not clear. Here, we developed a multiplexed spike variant protein microarray (SVPM) and utilized it for quantifying neutralizing activity, drug screening, and profiling humoral immunity. First, we demonstrated the competition between antispike antibody and ACE2 on SVPM for measuring the neutralizing activity against multiple spike variants. Next, we collected the serums from healthy subjects and COVID-19 patients with different severities and profile the neutralizing activity as well as antibody isotypes. We identified the inhibition of ACE2 binding was stronger against multiple variants in severe compared to mild/moderate or critical patients. Moreover, the serum IgG against nonstructural protein 3 was elevated in severe but not in mild/moderate and critical cases. Finally, we evaluated two ACE2 inhibitors, Ramipril and Perindopril, and found the dose-dependent inhibition of ACE2 binding to all the spike variants except for B.1.617.3. Together, the SVPM and the assay procedures provide a tool for profiling neutralizing antibodies, antibody isotypes, and reagent specificities.

. List of protein printed on the SVPM S3 Figure S2 . Heatmap visualization of ACE2 bindings against wild-type and common SARS-CoV-2 variants.

Serum from H, M, S, and C subjects were collected and analyzed for their ACE2 bindings by using SVPM. The gray color referred to outliers identified by Grubbs' method with alpha 0.0001. Figure S3 . Heatmap visualization of serum IgG against wild-type and common SARS-CoV-2 variants.

Serum from H, M, S, and C subjects were collected and analyzed for their IgG bindings by using SVPM. The gray color referred to outliers identified by Grubbs' method with alpha 0.0001. Figure S4 . Heatmap visualization of serum IgA against wild-type and common SARS-CoV-2 variants.

Serum from H, M, S, and C subjects were collected and analyzed for their IgA bindings by using SVPM. The gray color referred to outliers identified by Grubbs' method with alpha 0.0001. Figure S5 . Heatmap visualization of serum IgM against wild-type and common SARS-CoV-2 variants.

Serum from H, M, S, and C subjects were collected and analyzed for their IgM bindings by using SVPM. The gray color referred to outliers identified by Grubbs' method with alpha 0.0001.

Representative images from serum profiling by using the SVPM