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

Authors :: Viant C
Escolano A
Chen ST
Nussenzweig MC
Source :: STAR protocols [STAR Protoc] 2021 Mar 15; Vol. 2 (2), pp. 100389. Date of Electronic Publication: 2021 Mar 15 (Print Publication: 2021).
Publication Year :: 2021
Abstract: 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).<br />Competing Interests: M.C.N. is an inventor on the patent for 3BNC60. The rights to 3BNC60 have been licensed to Gilead by Rockefeller University.<br /> (© 2021 The Author(s).)

Subjects :: Animals
Female
Male
Mice
Protein Binding
Antibodies, Monoclonal analysis
Antibodies, Monoclonal chemistry
Antibodies, Monoclonal genetics
Antibodies, Monoclonal metabolism
Antigens analysis
Antigens chemistry
Antigens metabolism
B-Lymphocytes chemistry
Cloning, Molecular methods
Sequence Analysis, DNA methods
Single-Cell Analysis methods

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