Precise determination of the diversity of a combinatorial antibody library gives insight into the human immunoglobulin repertoire.

ABSTRACT: Antibody repertoire diversity, potentially as high as 10(11) unique molecules in a single individual, confounds characterization by conventional sequence analyses. In this study, we present a general method for assessing human antibody sequence diversity displayed on phage using massively parallel pyrosequencing, a novel application of Kabat column-labeled profile Hidden Markov Models, and translated complementarity determining region (CDR) capture-recapture analysis. Pyrosequencing of domain amplicon and RCA PCR products generated 1.5 x 10(6) reads, including more than 1.9 x 10(5) high quality, full-length sequences of antibody variable fragment (Fv) variable domains. Novel methods for germline and CDR classification and fine characterization of sequence diversity in the 6 CDRs are presen

SUBMITTER: Glanville J 

PROVIDER: S-EPMC2787155 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature