Laboratory and Data Analysis Methods for Characterization of Human B Cell Repertoires by High-Throughput DNA Sequencing

Methods Mol Biol. 2015:1343:219-33. doi: 10.1007/978-1-4939-2963-4_17.

Abstract

High-throughput DNA sequencing techniques have greatly accelerated the pace of research into the repertoires of antibody and T cell receptor gene rearrangements that confer antigen specificity to adaptive immune responses. Studies of aging-related changes in human B cell repertoires have benefited from the ability to detect and quantify thousands to millions of B cell clones in human samples, and study the mutational lineages and isotype switching relationships within each clonal lineage. Correlation of repertoire analysis with antibody gene data from antigen-specific B cells is poised to give much greater insight into clinically relevant B cell responses and memory storage. Here, we describe strategies for preparing and analyzing human antibody gene libraries for studying B cell repertoires.

Keywords: Aging; Antibody; Deep sequencing; High-throughput DNA sequencing; Immunoglobulin; Immunome; Repertoire.

MeSH terms

  • Antibodies / genetics
  • B-Lymphocytes / metabolism*
  • Computational Biology / methods
  • Gene Library
  • Gene Rearrangement, B-Lymphocyte
  • Genes, Immunoglobulin*
  • High-Throughput Nucleotide Sequencing*
  • Humans
  • Sequence Analysis, DNA

Substances

  • Antibodies