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Abstract for High-Throughput Compound Profiling Using Promega
Luminescent Assays on Tecan's EVO® 200 System
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.
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