HTRF Resource Library

Case Studies of uHTS compatible Detection Formats for Kinase Assays

By: T. Spicer

Scripps

Abstract

As part of our internal drug discovery efforts at Scripps Florida and also in conjunction with our participation in the NIH’s Molecular Libraries Screening Center Network (MLSCN), we have evaluated a variety of assay formats for kinases based on assay sensitivity, ease of application and economy. Three in-vitro enzymatic kinase assays were developed, miniaturized and implemented on a fully automated robotic screening system in 1536-well format utilizing three separate detection formats, from three different vendors. Of the three assays, one involves luminescence detection and the other two are based on time resolved fluorescence. All assays were found to be readily amenable to miniaturization and screening with reproducible results. However, the differences in detection specific to maintaining screening sensitivity for substrate and or ATP inhibitors/competitors can alter ones choice of detection. Case studies of the different formats will be presented.