Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells

The analysis of B cell receptors (BCR) from single B cells is crucial to understanding humoral immune responses. Here, we describe a protocol for the sequencing, cloning, and characterization of antibody genes that encode BCRs. We used this method to analyze the BCRs of different mouse B cell popula...

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Detalles Bibliográficos
Autores principales: Viant, Charlotte, Escolano, Amelia, Chen, Spencer T., Nussenzweig, Michel C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7985702/
https://www.ncbi.nlm.nih.gov/pubmed/33778783
http://dx.doi.org/10.1016/j.xpro.2021.100389
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author Viant, Charlotte
Escolano, Amelia
Chen, Spencer T.
Nussenzweig, Michel C.
author_facet Viant, Charlotte
Escolano, Amelia
Chen, Spencer T.
Nussenzweig, Michel C.
author_sort Viant, Charlotte
collection PubMed
description The analysis of B cell receptors (BCR) from single B cells is crucial to understanding humoral immune responses. Here, we describe a protocol for the sequencing, cloning, and characterization of antibody genes that encode BCRs. We used this method to analyze the BCRs of different mouse B cell populations for somatic hypermutations, clonal and phylogenic relationships, and their affinity for cognate antigen. For complete details on the use and execution of this protocol, please refer to Viant et al. (2020).
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spelling pubmed-79857022021-03-25 Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells Viant, Charlotte Escolano, Amelia Chen, Spencer T. Nussenzweig, Michel C. STAR Protoc Protocol The analysis of B cell receptors (BCR) from single B cells is crucial to understanding humoral immune responses. Here, we describe a protocol for the sequencing, cloning, and characterization of antibody genes that encode BCRs. We used this method to analyze the BCRs of different mouse B cell populations for somatic hypermutations, clonal and phylogenic relationships, and their affinity for cognate antigen. For complete details on the use and execution of this protocol, please refer to Viant et al. (2020). Elsevier 2021-03-15 /pmc/articles/PMC7985702/ /pubmed/33778783 http://dx.doi.org/10.1016/j.xpro.2021.100389 Text en © 2021 The Author(s) http://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
Viant, Charlotte
Escolano, Amelia
Chen, Spencer T.
Nussenzweig, Michel C.
Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells
title Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells
title_full Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells
title_fullStr Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells
title_full_unstemmed Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells
title_short Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells
title_sort sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse b cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7985702/
https://www.ncbi.nlm.nih.gov/pubmed/33778783
http://dx.doi.org/10.1016/j.xpro.2021.100389
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