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Analysis of Somatic Hypermutation in the JH4 intron of Germinal Center B cells from Mouse Peyer's Patches
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The Use of Somatic Hypermutation for the Affinity Maturation of Therapeutic Antibodies

Peter M Bowers1, William J Boyle2, Robert Damoiseaux3

  • 1Clinical and Translational Science Institute, UCLA David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA. pbowers@mednet.ucla.edu.

Methods in Molecular Biology (Clifton, N.J.)
|September 10, 2018
PubMed

The engineering of antibodies and antibody fragments for affinity maturation, stability, and other biophysical characteristics is a common aspect of therapeutic development. Maturation of antibodies in B cells during the adaptive immune response is the result of a process called somatic hypermutation (SHM), in which the activation-induced cytidine deaminase (AID) acts to introduce mutations into immunoglobulin (Ig) genes. Iterative selection and clonal expansion of B cells containing affinity-enhancing mutations drive an increase in the overall affinity of antibodies. Here we describe the use of SHM coupled with mammalian cell surface display for the maturation of antibodies in vitro and the complementarity of these methods with the mining of immune lineages using next-generation sequencing (NGS).

Keywords:
Affinity maturationAntibodyComplementarity-determining region (CDR)Fluorescence-activated cell sorting (FACS)Heavy chain (HC)Light chain (LC)Somatic hypermutation (SHM)

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