Quantitation of cytochrome P450 enzymes (CYP1A1/2, 2B11, 2C21 and 3A12) in dog liver microsomes by enzyme-linked immunosorbent assay

Abstract

1. An enzyme-linked immunosorbent assay (ELISA) using specific antisera has been developed to quantify individual cytochrome P450 (P450) enzymes (1A1/2, 2B11, 2C21 and 3A12) in dog liver microsomes.

2. The specific contents of CYP1A1/2, 2B11, 2C21 and 3A12 in untreated male dog liver microsomes determined by the ELISA were 17, 48, 160 and 69 pmol/mg protein respectively, corresponding to 4, 10, 34 and 15% of total optically determined P450 respectively. These P450 enzymes in untreated female dog liver microsomes showed almost similar amounts and relative proportions to those observed in male dog liver microsomes.

3. The oral treatment of male dogs with phenobarbital (PB), rifampicin (Rif) or β-naphthoflavone (β-NF) induced significant increases in the contents of CYP1A1/2 (12-fold by β-NF), 2B11 (16-fold by PB), 2C21 (2-fold by PB) and 3A12 (5-fold by PB and Rif), resulting in marked proportional alterations of the P450 enzymes in dog liver microsomes.

4. This ELISA method will be a useful tool for investigating possible influences (induction/suppression) of xenobiotics on the expression of P450 enzymes in dog liver.