View All Posters

P2034
A Surface Plasmon Resonance Based Assay for the Detection & Characterization of Promiscuous Inhibitors
Presenter , Anthony Giannetti, Roche Palo Alto, USA
Additional Authors: Bruce Koch, Michelle F. Browner 
Promiscuous binders achieve enzyme inhibition using a non-specific aggregation-type binding mechanism to proteins. These compounds are a source of false-positive hits in biochemical inhibition assays and should be removed from screening hit lists as they are not good candidates to initiate medicinal chemistry programs. We introduce a robust approach to identify these molecules early in the lead generation process using real time surface plasmon resonance based biosensors to observe the behavior of the binding interactions between promiscuous compounds and proteins. Furthermore, the time resolution of the assay reveals a number of distinct mechanisms that promiscuous compounds employ to inhibit enzyme function, and indicate that the type of mechanism can vary depending on the protein target. A classification scheme for these compounds is presented that can be used to rapidly characterize the hits from high-throughput screens and eliminate compounds with a non-specific mechanism of inhibition. For well behaved compounds the same assay also provides the binding constant, thereby providing additional data for hit prioritization.