Jason Quarles1, Tracy Worzella2, Brad Larson2, Jason Christiansen1, Ralph Ito1, Lorah Perlee1
1Protedyne Corporation, 1000 Day Hill Road, Windsor, CT 06095
2Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711
Many drug discovery programs incorporate high-throughput screening of cytochrome P450 activity to identify potential drug toxicities and improve the efficiency and cost-effectiveness of the drug development process. In this investigation, Promega P450-Glo™ kits for CYP1A1, CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 were evaluated on the Protedyne BioCube™ System, an industrial grade automation platform configured with a BMG Labtech PheraStar reader. Each enzyme was subject to the following tests performed in parallel manually and on the automated BioCube™ System: IC50 determinations, compound screening with known inhibitors, Z′-Factor, and limit of detection. IC50 determinations and compound screening exhibited good correlation between manually generated and BioCube™ System-generated inhibition values. Z′-Factor scores ranged from 0.69 to 0.79, revealing robust performance with good dynamic range of the automated assay. Limit of detection values ranged from 0.97–7.8fmol of enzyme, highlighting the sensitivity of the assays.