CYP Phenotyping, Recombinant CYP Isoforms, with Relative CYP Contribution

Assay System: 

  • Individually expressed recombinant human CYP isoforms (rhCYPs), including CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4.
  • Human CYP isoforms responsible for metabolism of test compound determined by measuring metabolic intrinsic clearance (CLint) in the presence of individual rhCYPs.

Assay Conditions: 

Test Compound Concentration 1 µM
Incubation Time 0, 5, 10, 15, 30, and 45 minutes
pH 7.4
Temperature 37°C
Analysis LC-MS/MS
Number of Repeats 2

Study Report: Contribution of individual rhCYPs to overall metabolism of test compound (in %) calculated from measured CLint values based on abundance of respective CYPs in liver and corresponding intersystem extrapolation factors (ISEF).1

Test Compound Requirement: 1-2 mg dry powder or 100-200 µL of 10 mM DMSO stock solution.

1 Chen, Y., Liu, L., Nguyen, K., Fretland, A.J. Utility of Intersystem Extrapolation Factors in Early Reaction Phenotyping and the Quantitative Extrapolation of Human Liver Microsomal Intrinsic Clearance Using Recombinant Cytochromes P450. Drug Metabolism and Disposition 39, 373-382 (2011).