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

doi:10.1002/cpim.116

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

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88 Scopus citations

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 language	English (US)
Article number	e116
Journal	Current Protocols in Immunology
Volume	131
Issue number	1
DOIs	https://doi.org/10.1002/cpim.116
State	Published - Dec 2020

Keywords

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

ASJC Scopus subject areas

Immunology

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10.1002/cpim.116

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Vanderheiden, A., Edara, V. V., Floyd, K., Kauffman, R. C., Mantus, G., Anderson, E., Rouphael, N., Edupuganti, S., Shi, P. Y., Menachery, V. D., Wrammert, J., & Suthar, M. S. (2020). Development of a Rapid Focus Reduction Neutralization Test Assay for Measuring SARS-CoV-2 Neutralizing Antibodies. Current Protocols in Immunology, 131(1), Article e116. https://doi.org/10.1002/cpim.116

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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.",

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AU - Mantus, Grace

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AU - Rouphael, Nadine

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AB - 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.

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

Abstract

Keywords

ASJC Scopus subject areas

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