Journal
PROTEIN ENGINEERING DESIGN & SELECTION
Volume 22, Issue 3, Pages 159-168Publisher
OXFORD UNIV PRESS
DOI: 10.1093/protein/gzn058
Keywords
antibody; human; library; phage display; scFv
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A large 1.29 x 10(11) antibody fragment library, based upon variable (V) genes isolated from human B-cells from 160 donors has been constructed and its performance measured against a panel of 28 different clinically relevant antigens. Over 5000 different target-specific antibodies were isolated to the 28 antigens with 3340 identified as modulating the biological function (e.g. antagonism, agonism) of the target antigen. This represents an average of similar to 120 different functionally active antibodies per target. Analysis of a sample of > 800 antibodies from the unselected library indicates V gene usage is representative of the human immune system with no strong bias towards any particular V-H-V-L pairing. Germline diversity is broad with 45/49 functional V-H germlines and 28/30 V-lambda and 30/35 V-kappa light-chain germlines represented in the sample. The number of functional V-H germlines and V-kappa light-chain germlines present is increased to 48/49 and 31/35, respectively, when selected V gene usage is included in the analysis. However, following selection on the antigen panel, V(H)1-V(lambda)1 germline family pairings are preferentially enriched and represent a remarkable 25% of the antigen-specific selected repertoire.
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