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A SARS-CoV-2 serological assay to determine the presence of blocking antibodies that compete for human ACE2 binding
As SARS-CoV-2 continues to spread around the world, there is an urgent need for new assay formats to characterize the humoral response to infection. Convalescent serum is being used for treatment and for isolation of patient-derived antibodies. However, currently there is not a simple means to estim...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7273274/ https://www.ncbi.nlm.nih.gov/pubmed/32511506 http://dx.doi.org/10.1101/2020.05.27.20114652 |
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author | Byrnes, James R. Zhou, Xin X. Lui, Irene Elledge, Susanna K. Glasgow, Jeff E. Lim, Shion A. Loudermilk, Rita Chiu, Charles Y. Wilson, Michael R. Leung, Kevin K. Wells, James A. |
author_facet | Byrnes, James R. Zhou, Xin X. Lui, Irene Elledge, Susanna K. Glasgow, Jeff E. Lim, Shion A. Loudermilk, Rita Chiu, Charles Y. Wilson, Michael R. Leung, Kevin K. Wells, James A. |
author_sort | Byrnes, James R. |
collection | PubMed |
description | As SARS-CoV-2 continues to spread around the world, there is an urgent need for new assay formats to characterize the humoral response to infection. Convalescent serum is being used for treatment and for isolation of patient-derived antibodies. However, currently there is not a simple means to estimate serum bulk neutralizing capability. Here we present an efficient competitive serological assay that can simultaneously determine an individual’s seropositivity against the SARS-CoV-2 Spike protein and estimate the neutralizing capacity of anti-Spike antibodies to block interaction with the human angiotensin converting enzyme 2 (ACE2) required for viral entry. In this ELISA-based assay, we present natively-folded viral Spike protein receptor binding domain (RBD)-containing antigens via avidin-biotin interactions. Sera are then supplemented with soluble ACE2-Fc to compete for RBD-binding serum antibodies, and antibody binding quantified. Comparison of signal from untreated serum and ACE2-Fc-treated serum reveals the presence of antibodies that compete with ACE2 for RBD binding, as evidenced by loss of signal with ACE2-Fc treatment. In our test cohort of nine convalescent SARS-CoV-2 patients, we found all patients had developed anti-RBD antibodies targeting the epitope responsible for ACE2 engagement. This assay provides a simple and high-throughput method to screen patient sera for potentially neutralizing anti-Spike antibodies to enable identification of candidate sera for therapeutic use. |
format | Online Article Text |
id | pubmed-7273274 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-72732742020-06-07 A SARS-CoV-2 serological assay to determine the presence of blocking antibodies that compete for human ACE2 binding Byrnes, James R. Zhou, Xin X. Lui, Irene Elledge, Susanna K. Glasgow, Jeff E. Lim, Shion A. Loudermilk, Rita Chiu, Charles Y. Wilson, Michael R. Leung, Kevin K. Wells, James A. medRxiv Article As SARS-CoV-2 continues to spread around the world, there is an urgent need for new assay formats to characterize the humoral response to infection. Convalescent serum is being used for treatment and for isolation of patient-derived antibodies. However, currently there is not a simple means to estimate serum bulk neutralizing capability. Here we present an efficient competitive serological assay that can simultaneously determine an individual’s seropositivity against the SARS-CoV-2 Spike protein and estimate the neutralizing capacity of anti-Spike antibodies to block interaction with the human angiotensin converting enzyme 2 (ACE2) required for viral entry. In this ELISA-based assay, we present natively-folded viral Spike protein receptor binding domain (RBD)-containing antigens via avidin-biotin interactions. Sera are then supplemented with soluble ACE2-Fc to compete for RBD-binding serum antibodies, and antibody binding quantified. Comparison of signal from untreated serum and ACE2-Fc-treated serum reveals the presence of antibodies that compete with ACE2 for RBD binding, as evidenced by loss of signal with ACE2-Fc treatment. In our test cohort of nine convalescent SARS-CoV-2 patients, we found all patients had developed anti-RBD antibodies targeting the epitope responsible for ACE2 engagement. This assay provides a simple and high-throughput method to screen patient sera for potentially neutralizing anti-Spike antibodies to enable identification of candidate sera for therapeutic use. Cold Spring Harbor Laboratory 2020-05-29 /pmc/articles/PMC7273274/ /pubmed/32511506 http://dx.doi.org/10.1101/2020.05.27.20114652 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Byrnes, James R. Zhou, Xin X. Lui, Irene Elledge, Susanna K. Glasgow, Jeff E. Lim, Shion A. Loudermilk, Rita Chiu, Charles Y. Wilson, Michael R. Leung, Kevin K. Wells, James A. A SARS-CoV-2 serological assay to determine the presence of blocking antibodies that compete for human ACE2 binding |
title | A SARS-CoV-2 serological assay to determine the presence of blocking antibodies that compete for human ACE2 binding |
title_full | A SARS-CoV-2 serological assay to determine the presence of blocking antibodies that compete for human ACE2 binding |
title_fullStr | A SARS-CoV-2 serological assay to determine the presence of blocking antibodies that compete for human ACE2 binding |
title_full_unstemmed | A SARS-CoV-2 serological assay to determine the presence of blocking antibodies that compete for human ACE2 binding |
title_short | A SARS-CoV-2 serological assay to determine the presence of blocking antibodies that compete for human ACE2 binding |
title_sort | sars-cov-2 serological assay to determine the presence of blocking antibodies that compete for human ace2 binding |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7273274/ https://www.ncbi.nlm.nih.gov/pubmed/32511506 http://dx.doi.org/10.1101/2020.05.27.20114652 |
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