Promega Corporation

Using Inhibitors to Improve Specificity in Cell-Based Luminogenic Caspase Assays...

Using Inhibitors to Improve Specificity in Cell-Based Luminogenic Caspase Assays Scientific Poster




Martha O’Brien, Rich Moravec, Mike Scurria1, Laurent Bernad1, Cass Brouette, Terry Riss, and Bob Bulleit
Promega Corporation, Madison, WI 53711 and 1Promega Biosciences, San Luis Obispo, CA 93401

We demonstrate the use of Promega 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. IC or EC 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 or optimization.

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  • Part# PS025
  • Printed in USA.

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