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

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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Main Authors: Rouers, Angeline, Tay, Matthew Zirui, Ng, Lisa F. P., Renia, Laurent
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2023
Subjects:
Science::Biological sciences
Antibody
Cell Isolation
Online Access:https://hdl.handle.net/10356/168926
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-1689262023-06-25T15:37:48Z 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 School of Biological Sciences Lee Kong Chian School of Medicine (LKCMedicine) Infectious Diseases Labs, A*STAR Science::Biological sciences Antibody Cell Isolation 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. Agency for Science, Technology and Research (A*STAR) National Medical Research Council (NMRC) Published version This work was supported by the Biomedical Research Council (BMRC); A*CRUSE (Vaccine monitoring project); the A*ccelerate GAP-funded project (ACCL/19-GAP064-R20H-H) from Agency for Science, Technology and Research (A*STAR); Singapore National Medical Research Council COVID-19 Research Fund (COVID19RF-001, COVID19RF-007, COVID19RF-008, COVID19RF0009, COVID19RF-0011, COVID19RF-060, OF-LCG19May-0034); US Food and Drug Administration (#75F40120C00085); and A*STAR COVID-19 Research funding (H/20/04/g1/006). 2023-06-22T07:35:50Z 2023-06-22T07:35:50Z 2023 Journal Article Rouers, A., Tay, M. Z., Ng, L. F. P. & Renia, L. (2023). B-cell ELISpot assay to analyze human memory B cell and plasmablast responses specific to SARS-CoV-2 receptor-binding domain. STAR Protocols, 4(1), 102130-. https://dx.doi.org/10.1016/j.xpro.2023.102130 2666-1667 https://hdl.handle.net/10356/168926 10.1016/j.xpro.2023.102130 36853725 2-s2.0-85148745845 1 4 102130 en ACCL/19-GAP064-R20H-H COVID19RF-001 COVID19RF-007 COVID19RF-008 COVID19RF0009 COVID19RF-0011 COVID19RF-060 OF-LCG19May-0034 H/20/04/g1/006 STAR Protocols © 2023 The Authors. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Science::Biological sciences
Antibody
Cell Isolation
spellingShingle Science::Biological sciences
Antibody
Cell Isolation
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
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.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Rouers, Angeline
Tay, Matthew Zirui
Ng, Lisa F. P.
Renia, Laurent
format Article
author Rouers, Angeline
Tay, Matthew Zirui
Ng, Lisa F. P.
Renia, Laurent
author_sort Rouers, Angeline
title 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_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_sort b-cell elispot assay to analyze human memory b cell and plasmablast responses specific to sars-cov-2 receptor-binding domain
publishDate 2023
url https://hdl.handle.net/10356/168926
_version_ 1772827732992851968

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