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Curr Protoc Immunol . Development of a Rapid Focus Reduction Neutralization Test Assay for Measuring SARS-CoV-2 Neutralizing Antibodies

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  • Curr Protoc Immunol . Development of a Rapid Focus Reduction Neutralization Test Assay for Measuring SARS-CoV-2 Neutralizing Antibodies


    Curr Protoc Immunol


    . 2020 Dec;131(1):e116.
    doi: 10.1002/cpim.116.
    Development of a Rapid Focus Reduction Neutralization Test Assay for Measuring SARS-CoV-2 Neutralizing Antibodies


    Abigail Vanderheiden 1 2 3 , Venkata Viswanadh Edara 1 2 3 , Katharine Floyd 1 2 3 , Robert C Kauffman 2 4 , Grace Mantus 2 4 , Evan Anderson 1 , Nadine Rouphael 2 5 , Sri Edupuganti 2 5 , Pei-Yong Shi 6 , Vineet D Menachery 7 , Jens Wrammert 2 4 , Mehul S Suthar 1 2 3



    Affiliations

    Abstract

    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 Basic Protocol: Focus reduction neutralization test Alternate Protocol: mNeonGreen-based focus reduction neutralization test (FRNT-mNG).

    Keywords: SARS-Cov-2; antibody neutralization; high-throughput neutralization assay; neutralizing antibodies; serological assay.

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