An Efficient Method to Generate Monoclonal Antibodies from Human B Cells
- PMID: 30539468
- DOI: 10.1007/978-1-4939-8958-4_5
An Efficient Method to Generate Monoclonal Antibodies from Human B Cells
Abstract
In the age of personalized medicine, an efficient method to generate monoclonal antibodies (mAbs) is essential for biomedical and immunotherapeutic research. Numerous aspects of basic B-cell biology can be studied at the monoclonal level, including B-cell development, antibody responses to infection or vaccination, and autoimmune responses. Single-cell B-cell receptor cloning allows for the rapid generation of antigen-specific mAbs in a matter of several weeks. In this chapter, we provide an efficient method to generate mAbs from peripheral blood plasmablasts and memory B cells induced by infection and vaccination. Additionally, we provide a protocol on how to optimize single-cell B-cell sorting for both single-cell B-cell receptor cloning and single-cell RNA-sequencing, for the application of studying B-cell specificity and function (spec-seq). This protocol can be easily adapted for other B-cell populations, B cells in tissues, and B cells from other organisms.
Keywords: B-cell receptor; Cloning; Humoral immunity; Infection; Memory B-cell; Monoclonal antibody; Plasmablast; Single-cell RNA-sequencing; Spec-seq; Vaccination.
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