4.7 Article

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

Journal

JCI INSIGHT
Volume 6, Issue 16, Pages -

Publisher

AMER SOC CLINICAL INVESTIGATION INC
DOI: 10.1172/jci.insight.150012

Keywords

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Funding

  1. Victorian Government and Medical Research Future Fund [GNT2002073]
  2. Emergent Ventures Fast Grant
  3. Paul Ramsay Foundation
  4. National Health and Medical Research Council (NHMRC) fellowships [208693/Z/17/Z]
  5. Jack Ma Foundation
  6. NHMRC Investigator grant
  7. Australian Research Council Discovery Early Career Researcher Award fellowship
  8. Australian Government Department of Health
  9. Australian Government NHMRC Institutes Infrastructure Support Scheme

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This study introduces a new rapid multiplex assay for measuring SARS-CoV-2 antibodies that can evaluate multiple RBD natural variants, filling a major gap in SARS-CoV-2 research and providing a method for selecting complementary monoclonal antibody candidates and rapidly identifying immune escape to emerging RBD variants following vaccination or natural infection.
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 angiotensinconverting 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.

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