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Application of phage display to high throughput antibody generation and characterization

Darren J Schofield1, Anthony R Pope2, Veronica Clementel3, Jenny Buckell4, Susan DJ Chapple2, Kay F Clarke2, Jennie S Conquer5, Anna M Crofts2, Sandra RE Crowther2, Michael R Dyson6, Gillian Flack7, Gareth J Griffin2, Yvette Hooks2, William J Howat8, Anja Kolb-Kokocinski2, Susan Kunze3, Cecile D Martin9, Gareth L Maslen2, Joanne N Mitchell8, Maureen O'Sullivan10, Rajika L Perera9, Wendy Roake2, S Paul Shadbolt2, Karen J Vincent11, Anthony Warford12, Wendy E Wilson2, Jane Xie2, Joyce L Young3 and John McCafferty6*

Author Affiliations

1 Abcam Ltd, Cambridge Science Park, Cambridge CB4 0FW, UK

2 Wellcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridgeshire CB10 1HH, UK

3 Cambridge Antibody Technology, Granta Park, Cambridge CB21 6GH, UK

4 Cancer Research UK, Research Monoclonal Antibody Services, Lincoln's Inn Fields, London WC2A 3PX, UK

5 Molecular Products Ltd, Thaxted CM6 2LT, UK

6 Department of Biochemistry, University of Cambridge, Downing Site, Cambridge CB2 1QW, UK

7 Cellular Histopathology Department, Bedford Hospital NHS Trust, Bedford MK42 9DJ, UK

8 Cancer Research UK, Cambridge Research Institute, Cambridge CB2 0RE, UK

9 GlaxoSmithKline Medicines Research Center, Stevenage SG1 2NY, UK

10 Genzyme Therapeutics Ltd, Cambridge Science Park, Cambridge CB4 0WG, UK

11 Novartis Institutes for BioMedical Research, Discovery Technologies, CH-4002 Basel, Switzerland

12 Astra Zeneca Innovation Centre China, Shanghai, 200041, China

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Genome Biology 2007, 8:R254  doi:10.1186/gb-2007-8-11-r254

Published: 29 November 2007

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.