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Simultaneous evaluation of antibodies that inhibit SARS-CoV-2 variants via multiplex assay
Ester Lopez, … , Wai-Hong Tham, Amy W. Chung
Ester Lopez, … , Wai-Hong Tham, Amy W. Chung
Published July 12, 2021
Citation Information: JCI Insight. 2021;6(16):e150012. https://doi.org/10.1172/jci.insight.150012.
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Resource and Technical Advance COVID-19

Simultaneous evaluation of antibodies that inhibit SARS-CoV-2 variants via multiplex assay

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Abstract

The SARS-CoV-2 receptor binding domain (RBD) is both the principal target of neutralizing antibodies and one of the most rapidly evolving domains, which can result in the emergence of immune escape mutations, limiting the effectiveness of vaccines and antibody therapeutics. To facilitate surveillance, we developed a rapid, high-throughput, multiplex assay able to assess the inhibitory response of antibodies to 24 RBD natural variants simultaneously. We demonstrate how this assay can be implemented as a rapid surrogate assay for functional cell-based serological methods to measure the SARS-CoV-2 neutralizing capacity of antibodies at the angiotensin-converting enzyme 2–RBD (ACE2-RBD) interface. We describe the enhanced affinity of RBD variants N439K, S477N, Q493L, S494P, and N501Y to the ACE2 receptor and demonstrate the ability of this assay to bridge a major gap for SARS-CoV-2 research, informing selection of complementary monoclonal antibody candidates and the rapid identification of immune escape to emerging RBD variants following vaccination or natural infection.

Authors

Ester Lopez, Ebene R. Haycroft, Amy Adair, Francesca L. Mordant, Matthew T. O’Neill, Phillip Pymm, Samuel J. Redmond, Wen Shi Lee, Nicholas A. Gherardin, Adam K. Wheatley, Jennifer A. Juno, Kevin J. Selva, Samantha K. Davis, Samantha L. Grimley, Leigh Harty, Damian F.J. Purcell, Kanta Subbarao, Dale I. Godfrey, Stephen J. Kent, Wai-Hong Tham, Amy W. Chung

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Figure 2

Validation of the SARS-CoV-2 inhibition multiplex assay.

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Validation of the SARS-CoV-2 inhibition multiplex assay.
(A) Plasma samp...
(A) Plasma samples from patients recovered from SARS-CoV-2 infection (n = 39) and SARS-CoV-2–negative healthy controls (n = 7) were tested for inhibition of the RBD-ACE2 interaction and (B) the S1-ACE2 interaction. SARS-CoV-2–positive subjects are colored based on COVID-19 illness severity. Orange squares, severe; yellow triangles, moderate; green inverted triangles, mild; gray, SARS-CoV-2–negative controls. COVID-19 subjects who tested negative on the multiplex assay are indicated by hollow symbols. (C) Spearman’s correlation between relative S1 and RBD IC50 percentage inhibition values. (D) Spearman’s correlation of the percentage ACE2 inhibition at a 1 in 100 dilution of plasma to RBD and the relative IC50 value. (E) Spearman’s correlation of multiplex ACE-RBD inhibition assay IC50 values and the titer obtained with the virus microneutralization assay. (F) Spearman’s correlation of multiplex ACE-Spike (S1) inhibition assay IC50 values and the titer obtained with the virus microneutralization assay. (G) Spearman’s correlation of ACE-RBD inhibition and (H) RBD binding of control mAbs. (I) Percentage ACE2 inhibition of control mAbs. Cutoff values to establish the positive/negative thresholds are indicated by dotted lines.

Copyright © 2023 American Society for Clinical Investigation
ISSN 2379-3708

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