A conformation-selective monoclonal antibody against a small molecule-stabilised signalling-deficient form of TNF
Daniel Lightwood, Rebecca Munro, John Porter, David McMillan, Bruce Carrington, Alison Turner, Anthony Scott-Tucker, Elizabeth S Hickford, Antje Schmidt, David Fox, Alison Maloney, Tom Ceska, Tim Bourne, James P. O’Connell, Alastair D. G. Lawson
Abstract
We have recently described the development of a series of small-molecule inhibitors of human tumour necrosis factor (TNF) that stabilise an open, asymmetric, signalling-deficient form of the soluble TNF trimer. Here, we describe the generation, characterisation, and utility of a monoclonal antibody that selectively binds with high affinity to the asymmetric TNF trimer-small molecule complex. The antibody helps to define the molecular dynamics of the apo TNF trimer, reveals the mode of action and specificity of the small molecule inhibitors, acts as a chaperone in solving the human TNF-TNFR1 complex crystal structure, and facilitates the measurement of small molecule target occupancy in complex biological samples. We believe this work defines a role for monoclonal antibodies as tools to facilitate the discovery and development of small-molecule inhibitors of protein-protein interactions.