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

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

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Article numbere116
JournalCurrent Protocols in Immunology
Volume131
Issue number1
DOIs
StatePublished - Dec 2020

Keywords

  • antibody neutralization
  • high-throughput neutralization assay
  • neutralizing antibodies
  • SARS-Cov-2
  • serological assay

ASJC Scopus subject areas

  • Immunology

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