从单个小鼠 B 细胞中分离的单克隆抗体的测序、克隆和抗原结合分析。

Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA.

Howard Hughes Medical Institute (HHMI), The Rockefeller University, New York, NY 10065, USA.

STAR Protoc. 2021 Mar 15;2(2):100389. doi: 10.1016/j.xpro.2021.100389. eCollection 2021 Jun 18.

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).

单细胞 B 细胞受体（BCR）的分析对于理解体液免疫反应至关重要。在此，我们描述了一种用于测序、克隆和鉴定编码 BCR 的抗体基因的方法。我们使用该方法分析了不同小鼠 B 细胞群体的 BCR 以检测体细胞超突变、克隆和系统发育关系，以及它们与同源抗原的亲和力。如需详细了解本方案的使用和执行，请参考 Viant 等人（2020 年）的研究。