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Application of phage display to high throughput antibody generation and characterization
Authors:Darren J Schofield  Anthony R Pope  Veronica Clementel  Jenny Buckell  Susan DJ Chapple  Kay F Clarke  Jennie S Conquer  Anna M Crofts  Sandra RE Crowther  Michael R Dyson  Gillian Flack  Gareth J Griffin  Yvette Hooks  William J Howat  Anja Kolb-Kokocinski  Susan Kunze  Cecile D Martin  Gareth L Maslen  Joanne N Mitchell  Maureen O'Sullivan  Rajika L Perera  Wendy Roake  S Paul Shadbolt  Karen J Vincent  Anthony Warford  Wendy E Wilson  Jane Xie  Joyce L Young  John McCafferty
Institution:Abcam Ltd, Cambridge Science Park, Cambridge CB4 0FW, UK. Darren.Schofield@abcam.com
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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