Sequencing, cloning, and antigen binding analysis of monoclonal antibodies isolated from single mouse B cells
Charlotte Viant, Amelia Escolano, Spencer T. Chen, Michel C. Nussenzweig
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).
Topics & Concepts
Cloning (programming)B-cell receptorBiologyB cellAntigenMonoclonal antibodyAntibodybreakpoint cluster regionMolecular biologyComputational biologyGeneGeneticsComputer scienceProgramming languageT-cell and B-cell ImmunologyMonoclonal and Polyclonal Antibodies ResearchImmune Cell Function and Interaction