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