Mapping of discontinuous conformational epitopes by amide hydrogen/deuterium exchange mass spectrometry and computational docking

• DOI: 10.1002/jmr.1169
• Publication Date: 2012-03-08T16:28:22Z
• Authors: Deepangi Pandit; Steven J. Tuske; Stephen J Coales; Sook Yen E; Anita Liu; Jessica E. Lee; Jeffrey A. Morrow; Jennifer F. Nemeth; Yoshitomo Hamuro
• Keywords: Docking (animal); Hydrogen–deuterium exchange; Amide

Understanding antigen–antibody interactions at the sub‐molecular level is of particular interest for scientific, regulatory, and intellectual property reasons, especially with increasing demand monoclonal antibody therapeutic agents. Although various techniques are available determination an epitope, there no widely applicable, high‐resolution, reliable method available. Here, a combination approach using amide hydrogen/deuterium exchange coupled proteolysis mass spectrometry (HDX‐MS) computational docking was applied to investigate interactions. HDX‐MS medium‐resolution, medium‐throughput technology that can be epitope identification. First, epitopes cytochrome c –E8, IL‐13–CNTO607, IL‐17A–CAT‐2200 identified were compared those by X‐ray co‐crystal structures. The in good agreement high‐resolution crystallography. Second, data used as constraints docking. More specifically, non‐epitope residues antigen designated binding ineligible during This approach, termed HDX‐DOCK, gave more tightly clustered poses than stand‐alone all examined improved results significantly –E8 interaction. Copyright © 2012 John Wiley & Sons, Ltd.