tetano
Editor, Senior Moderator
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[SUP] 1 2 3 [/SUP], Venkata Viswanadh Edara[SUP] 1 2 3 [/SUP], Katharine Floyd[SUP] 1 2 3 [/SUP], Robert C Kauffman[SUP] 2 4 [/SUP], Grace Mantus[SUP] 2 4 [/SUP], Evan Anderson[SUP] 1 [/SUP], Nadine Rouphael[SUP] 2 5 [/SUP], Sri Edupuganti[SUP] 2 5 [/SUP], Pei-Yong Shi[SUP] 6 [/SUP], Vineet D Menachery[SUP] 7 [/SUP], Jens Wrammert[SUP] 2 4 [/SUP], Mehul S Suthar[SUP] 1 2 3 [/SUP]
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.
. 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[SUP] 1 2 3 [/SUP], Venkata Viswanadh Edara[SUP] 1 2 3 [/SUP], Katharine Floyd[SUP] 1 2 3 [/SUP], Robert C Kauffman[SUP] 2 4 [/SUP], Grace Mantus[SUP] 2 4 [/SUP], Evan Anderson[SUP] 1 [/SUP], Nadine Rouphael[SUP] 2 5 [/SUP], Sri Edupuganti[SUP] 2 5 [/SUP], Pei-Yong Shi[SUP] 6 [/SUP], Vineet D Menachery[SUP] 7 [/SUP], Jens Wrammert[SUP] 2 4 [/SUP], Mehul S Suthar[SUP] 1 2 3 [/SUP]
Affiliations
- PMID: 33215858
- DOI: 10.1002/cpim.116
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.