B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 receptor-binding domain

B-cell ELISpot is an extremely sensitive assay based on the secretion of antibodies by B cells that requires the differentiation of B cells into antibody-secreting cells. Here, we describe the procedure to analyze both plasmablast (PB) and memory B cell (MBC) responses specific to SARS-CoV-2 recepto...

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Detalles Bibliográficos
Autores principales: Rouers, Angeline, Tay, Matthew Zirui, Ng, Lisa F.P., Renia, Laurent
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9910018/
https://www.ncbi.nlm.nih.gov/pubmed/36853725
http://dx.doi.org/10.1016/j.xpro.2023.102130
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author Rouers, Angeline
Tay, Matthew Zirui
Ng, Lisa F.P.
Renia, Laurent
author_facet Rouers, Angeline
Tay, Matthew Zirui
Ng, Lisa F.P.
Renia, Laurent
author_sort Rouers, Angeline
collection PubMed
description B-cell ELISpot is an extremely sensitive assay based on the secretion of antibodies by B cells that requires the differentiation of B cells into antibody-secreting cells. Here, we describe the procedure to analyze both plasmablast (PB) and memory B cell (MBC) responses specific to SARS-CoV-2 receptor-binding domain (RBD) in the context of acute SARS-CoV-2 infection and vaccination. We detail steps for MBC stimulation, MBC and PB plating, detection, and counting of total IgG and RBD-specific spots. For complete details on the use and execution of this protocol, please refer to Tay et al. (2022).(1)
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spelling pubmed-99100182023-02-09 B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 receptor-binding domain Rouers, Angeline Tay, Matthew Zirui Ng, Lisa F.P. Renia, Laurent STAR Protoc Protocol B-cell ELISpot is an extremely sensitive assay based on the secretion of antibodies by B cells that requires the differentiation of B cells into antibody-secreting cells. Here, we describe the procedure to analyze both plasmablast (PB) and memory B cell (MBC) responses specific to SARS-CoV-2 receptor-binding domain (RBD) in the context of acute SARS-CoV-2 infection and vaccination. We detail steps for MBC stimulation, MBC and PB plating, detection, and counting of total IgG and RBD-specific spots. For complete details on the use and execution of this protocol, please refer to Tay et al. (2022).(1) Elsevier 2023-02-09 /pmc/articles/PMC9910018/ /pubmed/36853725 http://dx.doi.org/10.1016/j.xpro.2023.102130 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Rouers, Angeline
Tay, Matthew Zirui
Ng, Lisa F.P.
Renia, Laurent
B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 receptor-binding domain
title B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 receptor-binding domain
title_full B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 receptor-binding domain
title_fullStr B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 receptor-binding domain
title_full_unstemmed B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 receptor-binding domain
title_short B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 receptor-binding domain
title_sort b-cell elispot assay to analyze human memory b cell and plasmablast responses specific to sars-cov-2 receptor-binding domain
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9910018/
https://www.ncbi.nlm.nih.gov/pubmed/36853725
http://dx.doi.org/10.1016/j.xpro.2023.102130
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