title: Laboratory and Data Analysis Methods for Characterization of Human B Cell Repertoires by High-Throughput DNA Sequencing.
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.
authors.affiliation: Department of Pathology, Stanford University, 300 Pasteur Drive, Stanford, CA, 94305, USA.
authors.forename: Chen
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Pathology, Stanford University, 300 Pasteur Drive, Stanford, CA, 94305, USA.|Biomedical Informatics Training Program, Stanford University, Stanford, CA, 94305, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Pathology, Stanford University, 300 Pasteur Drive, Stanford, CA, 94305, USA.
authors.forename: Krishna M
authors.initials: KM
authors.lastname: Roskin
authors.affiliation: Department of Pathology, Stanford University, 300 Pasteur Drive, Stanford, CA, 94305, USA.
authors.forename: Katherine J L
authors.initials: KJ
authors.lastname: Jackson
authors.affiliation: Department of Pathology, Stanford University, 300 Pasteur Drive, Stanford, CA, 94305, USA. sboyd1@stanford.edu.
authors.forename: Scott D
authors.initials: SD
authors.lastname: Boyd
mesh_terms: D000906:Antibodies / Q000235:genetics; D001402:B-Lymphocytes / Q000378:metabolism*; D019295:Computational Biology / Q000379:methods; D015723:Gene Library; D015322:Gene Rearrangement, B-Lymphocyte; D005803:Genes, Immunoglobulin; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D017422:Sequence Analysis, DNA
chemical_list: D000906:Antibodies
keywords: Aging; Antibody; Deep sequencing; High-throughput DNA sequencing; Immunoglobulin; Immunome; Repertoire