Development of a Rapid Focus Reduction Neutralization Test Assay for Measuring SARS‐CoV‐2 Neutralizing Antibodies

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  • Abigail Vanderheiden
    Department of Pediatrics, Division of Infectious Disease Emory University School of Medicine Atlanta Georgia
  • Venkata Viswanadh Edara
    Department of Pediatrics, Division of Infectious Disease Emory University School of Medicine Atlanta Georgia
  • Katharine Floyd
    Department of Pediatrics, Division of Infectious Disease Emory University School of Medicine Atlanta Georgia
  • Robert C. Kauffman
    Emory Vaccine Center Emory University School of Medicine Atlanta Georgia
  • Grace Mantus
    Emory Vaccine Center Emory University School of Medicine Atlanta Georgia
  • Evan Anderson
    Department of Pediatrics, Division of Infectious Disease Emory University School of Medicine Atlanta Georgia
  • Nadine Rouphael
    Emory Vaccine Center Emory University School of Medicine Atlanta Georgia
  • Sri Edupuganti
    Emory Vaccine Center Emory University School of Medicine Atlanta Georgia
  • Pei‐Yong Shi
    Department of Biochemistry and Molecular Biology The University of Texas Medical Branch Galveston Texas
  • Vineet D. Menachery
    Department of Microbiology and Immunology, Institute for Human Infection and Immunity, World Reference Center for Emerging Viruses and Arboviruses University of Texas Medical Branch Galveston Texas
  • Jens Wrammert
    Emory Vaccine Center Emory University School of Medicine Atlanta Georgia
  • Mehul S. Suthar
    Department of Pediatrics, Division of Infectious Disease Emory University School of Medicine Atlanta Georgia

Description

<jats:title>Abstract</jats:title><jats:p>SARS‐CoV‐2 is a recently emerged human coronavirus that has escalated to a pandemic. There are currently no approved vaccines for SARS‐CoV‐2, which causes severe respiratory illness or death. Defining the antibody response to SARS‐CoV‐2 will be essential for understanding disease progression, long‐term immunity, and vaccine efficacy. Here we describe two methods for evaluating the neutralization capacity of SARS‐CoV‐2 antibodies. The basic protocol is a focus reduction neutralization test (FRNT), which involves immunostaining infected cells with a chromogen deposit readout. The alternate protocol is a modification of the FRNT that uses an infectious clone−derived SARS‐CoV‐2 virus expressing a fluorescent reporter. These protocols are adapted for use in a high‐throughput setting, and are compatible with large‐scale vaccine studies or clinical testing. © 2020 Wiley Periodicals LLC</jats:p><jats:p><jats:bold>Basic Protocol</jats:bold>: Focus reduction neutralization test</jats:p><jats:p><jats:bold>Alternate Protocol</jats:bold>: mNeonGreen‐based focus reduction neutralization test (FRNT‐mNG)</jats:p>

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