[DDC 2019] InCELL Pulse™: A Novel Cellular Target Engagement Assay Platform for Drug Discovery

[DDC 2019] InCELL Pulse™: A Novel Cellular Target Engagement Assay Platform for Drug Discovery
File Name/Number:
Drug Discovery Chemistry 2019

Year:
2019

It is well established that ligand binding can protect proteins from thermal denaturation in a cellular milieu; however, the broad application of this cellular target engagement concept to drug discovery has been hindered by a lack of simple platforms with sensitive quantitative readouts. Cell-based thermal stabilization assays are valuable methods for particular applications but can require target-specific antibodies for immuno-assay readouts, can be low-throughput for Western blot formats, and can be biased against multi-domain proteins. Here we describe InCELL Pulse, which uses Eurofins DiscoverX Enzyme Fragment Complementation (EFC) technology to overcome these limitations of current thermal stabilization assays. We have successfully applied InCELL Pulse to rapidly measure quantitative cellular target engagement potency values for inhibitors of diverse protein classes, including kinases, methyltransferases, and hydrolases. We further demonstrate broad utility for InCELL Pulse across the human kinome, where we have validated assays for kinases from seven of the eight phylogenetic groups.