Abdel-Moneim, Ahmed S. and Berre, Marie Le and Paulovčáková, Terézia and Verissimo, Carolina De Marco and Doyle, Sean and Dalton, John P. and Masterson, Claire and Martínez, Eduardo Ribes and Walsh, Laura and Gormley, Conor and Laffey, John G. and McNicholas, Bairbre and Simpkin, Andrew J. and Kilcoyne, Michelle (2023) A new multiplex SARS-CoV-2 antigen microarray showed correlation of IgG, IgA, and IgM antibodies from patients with COVID-19 disease severity and maintenance of relative IgA and IgM antigen binding over time. PLOS ONE, 18 (3). pp. 1-31. ISSN 1932-6203

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Abstract

Zoonotic spillover of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to humans in December 2019 caused the coronavirus disease 2019 (COVID-19) pandemic. Serological monitoring is critical for detailed understanding of individual immune responses to infection and protection to guide clinical therapeutic and vaccine strategies. We developed a high throughput multiplexed SARS-CoV-2 antigen microarray incorporating spike (S) and nucleocapsid protein (NP) and fragments expressed in various hosts which allowed simultaneous assessment of serum IgG, IgA, and IgM responses. Antigen glycosylation influenced antibody binding, with S glycosylation generally increasing and NP glycosylation decreasing binding. Purified antibody isotypes demonstrated a binding pattern and intensity different from the same isotype in whole serum, probably due to competition from the other isotypes present. Using purified antibody isotypes from naïve Irish COVID-19 patients, we correlated antibody isotype binding to different panels of antigens with disease severity, with binding to the S region S1 expressed in insect cells (S1 Sf21) significant for IgG, IgA, and IgM. Assessing longitudinal response for constant concentrations of purified antibody isotypes for a patient subset demonstrated that the relative proportion of antigen-specific IgGs decreased over time for severe disease, but the relative proportion of antigen-specific IgA binding remained at the same magnitude at 5 and 9 months post-first symptom onset. Further, the relative proportion of IgM binding decreased for S antigens but remained the same for NP antigens. This may support antigen-specific serum IgA and IgM playing a role in maintaining longer-term protection, important for developing and assessing vaccine strategies. Overall, these data demonstrate the multiplexed platform as a sensitive and useful platform for expanded humoral immunity studies, allowing detailed elucidation of antibody isotypes response against multiple antigens. This approach will be useful for monoclonal antibody therapeutic studies and screening of donor polyclonal antibodies for patient infusions.

Item Type:	Article
Keywords:	new multiplex; SARS-CoV-2 antigen; microarray; correlation; IgG; IgA; IgM; antibodies; patients; COVID-19; disease; severity; maintenance; relative IgA; antigen binding;
Academic Unit:	Faculty of Science and Engineering > Biology
Item ID:	17112
Identification Number:	https://doi.org/10.1371/journal.pone.0283537
Depositing User:	Dr. Sean Doyle
Date Deposited:	24 Apr 2023 11:36
Journal or Publication Title:	PLOS ONE
Publisher:	Public Library of Science
Refereed:	Yes
URI:	http://www.plosone.org/
Use Licence:	This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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