Assay development and hit finding

We perform assay development, library screening and hit finding for any new kinase target. Cloning and purification of the protein can be offered as well. The high throughput screening equipment with liquid handling robots is state of the art. The IMAP (Immobilized Metal Assay for Phosphochemicals) technology has been introduced as the preferred assay system. Yet, other assays with Fluorescence polarization and Transcreener Kinase Assay with Direct Immunodetection of ADP, and Homogenous Time Resolved Fluorescence assays (HTRF) can be conducted as well. Before entering a screening campaign, the assays are adapted and optimized according to standard protocols for the ideal substrate/kinase concentrations, buffers, K_m [ATP, substrate], time course and IC₅₀ values for reference compounds. Selectivity ranking based on the binding affinity of the compound to the various kinases will be performed.

The EVL has been compiled from kinase inhibitors, which are in clinical and preclinical stage and which have been examined to a great detail. Therefore, the EVL is an ideal hit finding tool for novel kinase targets. CVL/EVL compounds are tested in biochemical assays for activity against a kinase of interest. Hit identification from the EVL shall be followed by testing the related part of the Master Library or by testing the whole Master Library to find novel hits. The selected and active inhibitors of a particular kinase are applied in therapeutically relevant cellular assays, in order to confirm the role of the kinase activity in this biological model. Validated hits from the NCL are good starting points for extended biological screening and compound optimization.