Engineering the surface properties of a human monoclonal antibody prevents self-association and rapid clearance in vivo
van der Walle, CF
Ridderstad Wollberg, A
Nature Publishing Group
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Uncontrolled self-association is a major challenge in the exploitation of proteins as therapeutics. Here we describe the development of a structural proteomics approach to identify the amino acids responsible for aberrant self-association of monoclonal antibodies and the design of a variant with reduced aggregation and increased serum persistence in vivo. We show that the human monoclonal antibody, MEDI1912, selected against nerve growth factor binds with picomolar affinity, but undergoes reversible self-association and has a poor pharmacokinetic profile in both rat and cynomolgus monkeys. Using hydrogen/deuterium exchange and cross-linking-mass spectrometry we map the residues responsible for self-association of MEDI1912 and show that disruption of the self-interaction interface by three mutations enhances its biophysical properties and serum persistence, whilst maintaining high affinity and potency. Immunohistochemistry suggests that this is achieved via reduction of non-specific tissue binding. The strategy developed represents a powerful and generic approach to improve the properties of therapeutic proteins.
We would like to thank Mrs Amy Barker (University of Leeds) for her help with bioanalytical ultracentrifugation. We acknowledge financial support from the Wellcome Trust (equipment grant 090932/Z/09/Z for EM) and CLP funding 092896, the ERC ((FP7/2007–2013)/ERC grant agreement no. 32240 (M.G.I., N.A.R. and S.E.R.), and the Biotechnology and Biological Sciences Research Council (BBSRC) for providing funds for MS equipment (BB/E012558/1) and for funding P.W.A.D (BB/J011819/1). We thank Dominic Corkhill and in vivo team at MedImmune Cambridge, UK, and also Mary McFarlane and the Toxicology team. We thank the MedImmune Biologics Expression and Early Material Supply Teams for antibody protein production.
This is the author accepted manuscript. The final version is available from Nature Publishing Group via the DOI in this record.
Vol. 6, pp. 38644 - 38644