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Abstract for Automating Cytochrome P450 Screening Using Promega
P450-Glo™ CYP450 Assays and the Protedyne BioCube™ System
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. |
