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

Overview

Journal Curr Protoc Immunol

Specialty Allergy & Immunology

Date 2020 Nov 20

PMID 33215858

Citations 76

Authors

Abigail Vanderheiden

Venkata Viswanadh Edara

Katharine Floyd

Robert C Kauffman

Grace Mantus

Evan Anderson

Nadine Rouphael

Sri Edupuganti

Pei-Yong Shi

Vineet D Menachery

Jens Wrammert

Mehul S Suthar

Affiliations

Soon will be listed here.

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).

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