John McCafferty

Communications between cells are directed through surface receptors which are activated through contact with neighbouring cells or via secreted ligands from distant cells. Recombinant antibodies provide a means to control receptor activation with potential utility in the treatment of cancer or the control of stem cell differentiation. Using phage display we have isolated and improved the affinity of several human antibodies which block receptor signalling.

-  CXCL12 is a key mediator in recruiting fibroblasts, leukocytes and endothelial cells to provide a supportive microenvironment for tumour cells. We have generated human antibodies which block the interaction of CXCL12 with its receptor CXCR4 with the potential to act at multiple levels.

-  We have also made blocking antibodies to c-Met, the receptor for the cell motility factor HGF/SF. The key outcome of c-met signalling is the migration of cancer cells from the primary tumour to the local lymphatic or blood capillaries where they initiate the later stages of metastasis. C-met is therefore a key target for the therapy of invasive and metastatic cancer.

-   We have generated antibodies which block ADAM17, a cancer associated metalloprotease, in collaboration with Gill Murphy and Chris Tape at CRI (ref 1). Affinity matured variants are currently being tested in tumour xenograft models.

-   Finally we have generated antibodies which specifically block either Notch-1 or Notch-2 and are are investigating their potential in controlling stem cell differentiation.

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.

Lab members: Kothai Nachiar Devi, Mike Dyson, Anna Melidoni, Tony Pope, Peter Slavny, Aneesh Karatt Vellatt

Key publications:

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

2.  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

3.  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

4.  Schofield DJ, Pope A……, *McCafferty J *(2007)* *Application of phage display to high throughput antibody generation and characterisation/. /Genome Biology. 8 R254