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

Research output: Contribution to journal › Article › peer-review

82 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

Access to Document

10.1002/cpim.116

Cite this

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

@article{2d6619cf68b3450dadbff3655c8f2aa1,

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

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.",

keywords = "SARS-Cov-2, antibody neutralization, high-throughput neutralization assay, neutralizing antibodies, serological assay",

author = "Abigail Vanderheiden and Edara, {Venkata Viswanadh} and Katharine Floyd and Kauffman, {Robert C.} and Grace Mantus and Evan Anderson and Nadine Rouphael and Sri Edupuganti and Shi, {Pei Yong} and Menachery, {Vineet D.} and Jens Wrammert and Suthar, {Mehul S.}",

note = "Publisher Copyright: {\textcopyright} 2020 Wiley Periodicals LLC",

year = "2020",

month = dec,

doi = "10.1002/cpim.116",

language = "English (US)",

volume = "131",

journal = "Current Protocols in Immunology",

issn = "1934-3671",

publisher = "John Wiley and Sons Inc.",

number = "1",

}

TY - JOUR

T1 - Development of a Rapid Focus Reduction Neutralization Test Assay for Measuring SARS-CoV-2 Neutralizing Antibodies

AU - Vanderheiden, Abigail

AU - Edara, Venkata Viswanadh

AU - Floyd, Katharine

AU - Kauffman, Robert C.

AU - Mantus, Grace

AU - Anderson, Evan

AU - Rouphael, Nadine

AU - Edupuganti, Sri

AU - Shi, Pei Yong

AU - Menachery, Vineet D.

AU - Wrammert, Jens

AU - Suthar, Mehul S.

PY - 2020/12

Y1 - 2020/12

N2 - 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.

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.

KW - SARS-Cov-2

KW - antibody neutralization

KW - high-throughput neutralization assay

KW - neutralizing antibodies

KW - serological assay

UR - http://www.scopus.com/inward/record.url?scp=85096457649&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85096457649&partnerID=8YFLogxK

U2 - 10.1002/cpim.116

DO - 10.1002/cpim.116

M3 - Article

C2 - 33215858

AN - SCOPUS:85096457649

SN - 1934-3671

VL - 131

JO - Current Protocols in Immunology

JF - Current Protocols in Immunology

IS - 1

M1 - e116

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this