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Assessment of B cell repertoire in humans

Wu, Y-C, Kipling, D and Dunn-Walters, D (2015) Assessment of B cell repertoire in humans Methods in Molecular Biology, 1343. pp. 199-218.

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Abstract

The B cell receptor (BCR) repertoire is highly diverse. Repertoire diversity is achieved centrally by somatic recombination of immunoglobulin (Ig) genes and peripherally by somatic hypermutation and Ig heavy chain class-switching. Throughout these processes, there is selection for functional gene rearrangements, selection against gene combinations resulting in self-reactive BCRs, and selection for BCRs with high affinity for exogenous antigens after challenge. Hence, investigation of BCR repertoires from different groups of B cells can provide information on stages of B cell development and shed light on the etiology of B cell pathologies. In most instances, the third complementarity determining region of the Ig heavy chain (CDR-H3) contributes the majority of amino acids to the antibody/antigen binding interface. Although CDR-H3 spectratype analysis provides information on the overall diversity of BCR repertoires, this fairly simple technique analyzes the relative quantities of CDR-H3 regions of each size, within a range of approximately 10–80 bp, without sequence detail and thus is limited in scope. High-throughput sequencing (HTS) techniques on the Roche 454 GS FLX Titanium system, however, can generate a wide coverage of Ig sequences to provide more qualitative data such as V, D, and J usage as well as detailed CDR3 sequence information. Here we present protocols in detail for CDR-H3 spectratype analysis and HTS of human BCR repertoires.

Item Type: Article
Subjects : Biosciences and Medicine
Divisions : Faculty of Health and Medical Sciences > School of Biosciences and Medicine
Authors :
AuthorsEmailORCID
Wu, Y-CUNSPECIFIEDUNSPECIFIED
Kipling, DUNSPECIFIEDUNSPECIFIED
Dunn-Walters, DUNSPECIFIEDUNSPECIFIED
Date : 2015
Identification Number : 10.1007/978-1-4939-2963-4_16
Copyright Disclaimer : The final publication is available at Springer via http://dx.doi.org/10.1007/978-1-4939-2963-4_16
Uncontrolled Keywords : B cell B cell receptor Immunoglobulin Antibody Repertoire High-throughput sequencing Next generation sequencing Spectratyping
Depositing User : Symplectic Elements
Date Deposited : 06 Jul 2016 15:03
Last Modified : 06 Jul 2016 15:03
URI: http://epubs.surrey.ac.uk/id/eprint/811135

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