Standardized two-step testing of antibody activity in COVID-19 convalescent plasma
Pavlo Gilchuk,
Isaac Thomsen,
Sandra Yoder,
Eric Brady,
James D. Chappell,
Laura J. Stevens,
Mark R. Denison,
Rachel E. Sutton,
Rita E. Chen,
Laura A. VanBlargan,
Naveenchandra Suryadevara,
Seth J. Zost,
Jonathan Schmitz,
Jill M. Pulley,
Michael S. Diamond,
Jillian P. Rhoads,
Gordon R. Bernard,
Wesley H. Self,
Todd W. Rice,
Allison P. Wheeler,
James E. Crowe, Jr.,
Robert H. Carnahan
Affiliations
Pavlo Gilchuk
Vanderbilt Vaccine Center, Vanderbilt University Medical Center, 11475 Medical Research Building IV, 2213 Garland Avenue, Nashville, TN 37232-0417, USA
Isaac Thomsen
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA; Vanderbilt Vaccine Research Program, Vanderbilt University Medical Center, Nashville, TN, USA
Sandra Yoder
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA; Vanderbilt Vaccine Research Program, Vanderbilt University Medical Center, Nashville, TN, USA
Eric Brady
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA; Vanderbilt Vaccine Research Program, Vanderbilt University Medical Center, Nashville, TN, USA
James D. Chappell
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA
Laura J. Stevens
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA
Mark R. Denison
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USA
Rachel E. Sutton
Vanderbilt Vaccine Center, Vanderbilt University Medical Center, 11475 Medical Research Building IV, 2213 Garland Avenue, Nashville, TN 37232-0417, USA
Rita E. Chen
Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA; Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO, USA
Laura A. VanBlargan
Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA
Naveenchandra Suryadevara
Vanderbilt Vaccine Center, Vanderbilt University Medical Center, 11475 Medical Research Building IV, 2213 Garland Avenue, Nashville, TN 37232-0417, USA
Seth J. Zost
Vanderbilt Vaccine Center, Vanderbilt University Medical Center, 11475 Medical Research Building IV, 2213 Garland Avenue, Nashville, TN 37232-0417, USA
Jonathan Schmitz
Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USA; Department of Urology, Vanderbilt University Medical Center, Nashville, TN 37232, USA
Jill M. Pulley
Vanderbilt Institute for Clinical and Translational Research, Vanderbilt University Medical Center, Nashville, TN, USA
Michael S. Diamond
Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA; Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, MO, USA; Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO, USA; Andrew M. and Jane M. Bursky Center for Human Immunology and Immunotherapy Programs, Washington University School of Medicine, Saint Louis, MO, USA
Jillian P. Rhoads
Vanderbilt Institute for Clinical and Translational Research, Vanderbilt University Medical Center, Nashville, TN, USA
Gordon R. Bernard
Vanderbilt Institute for Clinical and Translational Research, Vanderbilt University Medical Center, Nashville, TN, USA; Department of Medicine, Division of Allergy, Pulmonary and Critical Care Medicine, Vanderbilt University Medical Center, Nashville, TN 37232, USA
Wesley H. Self
Vanderbilt Institute for Clinical and Translational Research, Vanderbilt University Medical Center, Nashville, TN, USA; Department of Emergency Medicine, Vanderbilt University Medical Center, Nashville, TN 37232, USA
Todd W. Rice
Vanderbilt Institute for Clinical and Translational Research, Vanderbilt University Medical Center, Nashville, TN, USA; Department of Medicine, Division of Allergy, Pulmonary and Critical Care Medicine, Vanderbilt University Medical Center, Nashville, TN 37232, USA
Allison P. Wheeler
Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USA
James E. Crowe, Jr.
Vanderbilt Vaccine Center, Vanderbilt University Medical Center, 11475 Medical Research Building IV, 2213 Garland Avenue, Nashville, TN 37232-0417, USA; Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA; Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232, USA
Robert H. Carnahan
Vanderbilt Vaccine Center, Vanderbilt University Medical Center, 11475 Medical Research Building IV, 2213 Garland Avenue, Nashville, TN 37232-0417, USA; Department of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232, USA; Corresponding author
Summary: The COVID-19 pandemic revealed an urgent need for rapid profiling of neutralizing antibody responses and development of antibody therapeutics. The current Food and Drug Administration-approved serological tests do not measure antibody-mediated viral neutralization, and there is a need for standardized quantitative neutralization assays. We report a high-throughput two-step profiling approach for identifying neutralizing convalescent plasma. Screening and downselection for serum antibody binding to the receptor-binding domain are followed by quantitative neutralization testing using a chimeric vesicular stomatitis virus expressing spike protein of SARS-CoV-2 in a real-time cell analysis assay. This approach enables a predictive screening process for identifying plasma units that neutralize SARS-CoV-2. To calibrate antibody neutralizing activity in serum from convalescent plasma donors, we introduce a neutralizing antibody standard reagent composed of two human antibodies that neutralize SARS-CoV strains, including SARS-CoV-2 variants of concern. Our results provide a framework for establishing a standardized assessment of antibody-based interventions against COVID-19.
