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

被引：6
|
作者：
Viant, Charlotte [1 ]
Escolano, Amelia [1 ]
Chen, Spencer T. [1 ]
Nussenzweig, Michel C. [1 ,2 ]
机构：
[1] Rockefeller Univ, Lab Mol Immunol, New York, NY 10065 USA
[2] Rockefeller Univ, Howard Hughes Med Inst HHMI, New York, NY 10065 USA
来源：
STAR PROTOCOLS | 2021年 / 2卷 / 02期
关键词：
Protocol describes; BCR analysis; somatic; relationships; antigen; MEMORY;
D O I：
10.1016/j.xpro.2021.100389
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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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页数：23
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