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John McCafferty

Research Interests

We generate monoclonal antibodies by phage display. The entire process of monoclonal antibody generation by phage display relies on simple molecular biology techniques and so is well suited for high-­throughput antibody development. We have generated a high quality antibody display library of over 10 billion clones and have used this to isolate antibodies to hundreds of targets (Schofield et al, 2007). In a recent international pilot study we demonstrated the potential for high throughput antibody generation by generating a large panel of antibodies to a family of 20 human signalling molecules (Pershad et al, 2009). Furthermore we have shown that affinity maturation can be carried out in parallel on multiple antigens, resulting in a scalable method to generate antibodies with high affinity and utility. The resulting antibodies were used to identify interacting partners of the signalling molecule Shc1 using immunoprecipitation and mass spectrometry.

Keywords

monoclonal antibodies ; phage display

Key Publications

Bradbury A, Duebel S, Sidhu D, McCafferty J. (2011) Beyond natural antibodies: the power of in vitro display technologies. Nature Biotechnology 29 245-254

Tape CJ, Willems SH, Dombernowsky SL, Stanley PL, Fogarasi M, Ouwehand W, McCafferty J, Murphy G. (2011) Cross-Domain Inhibition of TACE Ectodomain Proc Natl Acad Sci 108 5578-5583

Dyson MR, Zheng Y, Zhang C, Colwill K, Pershad K, Kay BK, Pawson T, McCafferty J. Mapping protein interactions by combining antibody affinity maturation and mass spectrometry. Anal Biochem. 2011 Oct 1;417(1):25-35.

Pershad K, Pavlovic JD, Gräslund S, Nilsson P, Colwill K, Vellatt AK, Schofield DJ, Dyson MR, Pawson T, Kay BK, McCafferty J (2010) Generating a panel of highly specific antibodies to 20 human SH2 domains by phage display. Protein Engineering, Design and Selection vol. 23 279–288

Schofield DJ, Pope AR, Clementel V, Buckell J, Chapple SDj, Clarke KF, Conquer JS, Crofts AM, Crowther SR, Dyson MR, Flack G, Griffin GJ, Hooks Y, Howat WJ, Kolb-Kokocinski A, Kunze S, Martin CD, Maslen GL, Mitchell JN, O'Sullivan M, Perera RL, Roake W, Shadbolt SP, Vincent KJ, Warford A, Wilson WE, Xie J, Young JL, McCafferty J. Application of phage display to high throughput antibody generation and characterisation Genome Biology 2007. 8 R254