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Application of phage display to high throughput antibody generation and characterization.


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Authors

Schofield, Darren J 
Pope, Anthony R 
Clementel, Veronica 
Buckell, Jenny 
Chapple, Susan Dj 

Abstract

We have created a high quality phage display library containing over 1010 human antibodies and describe its use in the generation of antibodies on an unprecedented scale. We have selected, screened and sequenced over 38,000 recombinant antibodies to 292 antigens, yielding over 7,200 unique clones. 4,400 antibodies were characterized by specificity testing and detailed sequence analysis and the data/clones are available online. Sensitive detection was demonstrated in a bead based flow cytometry assay. Furthermore, positive staining by immunohistochemistry on tissue microarrays was found for 37% (143/381) of antibodies. Thus, we have demonstrated the potential of and illuminated the issues associated with genome-wide monoclonal antibody generation.

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Keywords

Animals, Antibody Formation, Antibody Specificity, Bacteriophages, Base Sequence, DNA Primers, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Gene Expression Profiling, Humans, Immunohistochemistry, In Situ Hybridization, Mice, Recombinant Proteins

Journal Title

Genome Biol

Conference Name

Journal ISSN

1474-760X
1474-760X

Volume Title

Publisher

Springer Science and Business Media LLC