An efficient method to generate monoclonal antibodies from human B cells

JJ Guthmiller, HL Dugan, KE Neu, LYL Lan… - … Antibodies: Methods and …, 2019 - Springer
JJ Guthmiller, HL Dugan, KE Neu, LYL Lan, PC Wilson
Human Monoclonal Antibodies: Methods and Protocols, 2019Springer
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 …
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.
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