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Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells
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An Efficient Method to Generate Monoclonal Antibodies from Human B Cells

Jenna J Guthmiller1, Haley L Dugan2, Karlynn E Neu1,2

  • 1Department of Medicine, Section of Rheumatology, The Knapp Center for Lupus and Immunology, University of Chicago, Chicago, IL, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 13, 2018
PubMed

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 receptorCloningHumoral immunityInfectionMemory B-cellMonoclonal antibodyPlasmablastSingle-cell RNA-sequencingSpec-seqVaccination

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