Tracy Worzella1, Brad Larson1, John Watson1, and Siegfried Sasshofer2
1Promega Corporation, 2800 Woods Hollow Road, Madison WI 53711, USA 2TECAN Austria Ges.m.b.H. Untersbergstrasse 1A, A-5082 Groedig
We demonstrate the use of Promega’s luminescent HTS assays for profiling test compounds on a Tecan system. This profiling example takes a parallel approach to compound analysis by incorporating diverse assay types including cell-based assays for viability and apoptosis induction, a cell-based GPCR DRD1 assay, cytochrome P450, P-glycoprotein, monoamine oxidase,
and kinase assays. Using a panel of assays, we show that one can obtain a better understanding of drug compound properties in order to better predict off-target activity and toxicity.
For this application, we have chosen several kinase (PKA) inhibitors and demonstrate the vast amount of information that can be obtained from these compounds by assaying them against a variety of chemistries. To generate the data, a Tecan Freedom Evo® 200 with an integrated TeMo™ was used to dispense cells, serially dilute test compounds and assemble assays in 384-well format. Luminescence was recorded with a Tecan GENios Pro™ plate reader. IC50 or EC50 calculations were performed for each compound and assay combination. Results show that data from these assays can be used to determine multiple compound characteristics for subsequent lead selection.