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. Here we describe how to create your own cellular target engagement assays using InCELL Pulse, which uses DiscoverX Enzyme Fragment Complementation (EFC) technology to overcome 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.
What will be covered?
- Overview of compound-target engagement
- Tools needed to create your own target engagement assay
- Experimental design and workflow
- Applications of InCELL Pulse assays
Who should attend?
- Enzymologists studying kinases, hydrolases, methyltransferases, and other enzymes
- Researchers developing compound-target engagement assays
- Basic research and assay development drug discovery scientists