Primary hepatocytes from three separate donors are isolated and cultured for the CYP Induction assay. Then they are exposed to each test compound and control samples for 72 hours. The tested compounds are evaluated at several different concentrations based on the expected human plasma drug concentrations.
After exposure, the mRNA is isolated from the cells in order to study the gene expression levels of the CYP enzymes CYP1A2, CYP3A4, and CYP2B6. This analysis is done using real-time RT-PCR (qRT-PCR). The resulting data is normalized according to housekeeping genes and is then presented as the fold induction compared to the negative (vehicle) control sample.
IonTox can also provide you with data for the catalytic activities of the CYP enzymes and assess CYP activity changes in relation to specific substrates. This information gives you a more complete assessment and an increased level of confidence.
CYP Induction assays are indispensible in the pharmaceutical industry, as any unwanted drug-drug interactions can be very costly for a pharmaceutics company. These induction assays are also utilized for different industrial chemicals and other consumer products.
All of our CYP Induction assays are carried out per the specific FDA guidelines and regulations regarding Drug Interaction Studies. These include testing at several concentrations, utilizing multiple control samples, and mRNA expression of the CYP enzymes with qRT-PCR.