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Automation of Luminescence-based CYP Inhibition Assays using BioTek Instrumentation for use in Drug Discoveryダウンロード
April 29, 2009
Authors: Brad Larson, Peter Banks, Jason Greene, BioTek Instruments, Inc., Sarah Shultz, Jim Cali, Mary Sobol, Simon Allard, Promega Corporation
The ability to determine the effects that lead compounds have on cytochrome P450 enzymes is an important part of today’s drug discovery routine. One desired component of this process is the ability to profile compounds against multiple CYP450 enzymes using the same basic procedure. The second is easy to use, yet dependable instrumentation that can be used to dilute compounds, as well as dispense assay reagents, that will create accurate titration curves.
Here we demonstrate the automation of the profiling process, from compound titration through assay component transfer, using BioTek’s Precision™ XS Automated Pipetting System. IC50’s of small molecule drugs were determined using recombinant CYP isoforms 3A4, 2C9 and 2D6 and luminogenic substrates specific to each from Promega’s P450-Glo™ CYP450 Screening Systems. Compounds were profiled against all three isoforms on the same 384-well assay plate to demonstrate the ease of this combined procedure.