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Automated Multiplexed Cell-Based Cytochrome P450 AssaysDownload
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July 12, 2010
Authors: Brad Larson, Peter Banks, BioTek Instruments, Winooski, VT; James J Cali, Mary Sobol, Promega Corporation, Madison, WI; Timothy Moeller, Celsis In Vitro Technologies, Baltimore, MD
This poster was presented at Drug-Drug Interactions June 14-16, 2010.
Cytochrome P450 enzymes are key players in the metabolism of drugs within the body. Therefore, it is essential to understand how these enzymes can be affected by xenobiotics with regards to inhibition, induction, and toxicity to avoid potential drug-drug interactions. Using a multiplexed format, where multiple pieces of data are obtained from a single well, can facilitate this process and provide more reliable results. Here we examine data from two separate projects combining assays to examine CYP activity, as well as toxicity, in a cell-based format. The first demonstrates the ability to monitor induction and inhibition of the CYP3A4 enzyme, as well as cytotoxicity from a single assay well, using DPX-2 cells. The second demonstrates the ability to monitor CYP1A2 and 3A4 induction, combined with cytotoxicity measurements, from a single well using cryopreserved hepatocytes. Each assay utilized simple automation that can easily fit into existing biosafety cabinets. Data shown confirms how the combination of assay, cells, and instrumentation provide a simplified, robust method to obtain this valuable information.