Cytochrome P450 Assay Services

Cytochromes (CYPs) are hemeproteins that play key roles in the metabolism of drugs. Understanding a compound’s inhibitory activity against key CYP proteins is essential for predicting drug-drug interactions. Reaction Biology provides compound profiling against the 6 most important CYP isoforms to provide early guidance on a compound's toxicity.

  • Low and large scale screening, as well as high-throughput screening options, are available
  • Custom-tailored assay development possible
  • The inhibition of CYP activity by drug candidates is measured via fluorescence readout

List of Assays

Name Synonym Data
CYP 1A2 CYP1A2 Data sheet
CYP 2C19 CYP2C19 Data sheet
CYP 2C9 CYP2C9 Data sheet
CYP 2D6 CYP2D6 Data sheet
CYP 3A4 CYP3A4 Data sheet
CYP 19A Aromatase, CYP19A1 Data sheet

Assay Details

Assay Format

For screening of CYP450, we use the Vivid CYP450 Screening Kits (by ThermoFisher) which enables the rapid measurement of interactions between drug candidates and cytochrome P450 enzymes using a fluorescent assay. Test compounds are analyzed by their capacity to inhibit the production of a fluorescent signal in reactions using CYP-containing baculosomes and CYP-specific substrates. Baculosomes are microsomes prepared from insect cells infected with recombinant baculovirus expressing a human CYP450 isoform.

CYP19A activity is measured with the Aromatase (CYP19A) Inhibitor Screening Kit (BioVision) which is a fluorescence-based assay. The assay uses recombinant human CYP19A and a fluorogenic substrate resulting in a fluorescent metabolite that can be quantified by a standard fluorescence reader.

Assay Setup

Setups: Single-dose screening in duplicates or IC50 value determination with 5 or 10 concentrations. Other screening formats are available upon request.

Controls: No inhibitor (DMSO vehicle) control and for every assay, one target-specific control compound is tested in 10-dose IC50 format. 

Turnaround time: 10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to the commencement of the studies.

Report: The raw data, % enzyme activity and control compound IC50 values will be reported in Excel format for single-dose assays. For IC50 orders, raw data, IC50 values, and curve fitting will be delivered in Excel format. Assay conditions, target, and substrate information are available upon request. Requirements for this information should be noted prior to the commencement of the study.

Screening facility: This assay is performed at our screening facility in Malvern, PA, US.

Compound requirements: In brief, for a standard project, 20 µl of a 10 mM DMSO stock or solid material is needed. Less material is needed for large scale screening. Please refer to our FAQs for information regarding compound preparation and shipping.