An inhibitory monoclonal antibody to human cytochrome P450 that specifically binds and inhibits P4502C9II, an allelic variant of P4502C9 having a single amino acid change Arg144 Cys.
A monoclonal antibody (MAb 292-2-3) has been isolated that binds specifically to a single allele of three expressed human cytochrome P4502C9 alleles. The MAb binds to 2C9Cys144 (II), and does not bind to the wild-type 2C9Arg144 (I), or the third allele 2C9Ile-->Leu359 (III) and thus the MAb detects an allele with > 99% homology and differing from the wild-type 2C9Arg144 (I) by a single amino acid. The MAb 292-2-3 does not bind to the other 2C isoforms (2C8, 2C18, 2C19) or the other human cytochrome P450s, 1A1, 1A2, 2A6, 2B6, 2C8, 2D6, 2E1 or 3A4/5. MAb 292-2-3 inhibits the metabolism of tolbutamide, diclofenac and phenanthrene by the target 2C9Cys144 (II) allele by > 90% and does not inhibit the catalytic activity of the wild-type 2C9Arg144 (I), or 2C9Ile-->Leu359 (III) the other 2C isoforms 2C8, 2C18, 2C19, or the other non-2C human P450s listed above. The MAb 292-2-3 is thus a prototype of an ideal and extraordinarily specific reagent for the detection and measurement of the metabolic role of highly related isoforms and polymorphic alleles of human cytochrome P450s. MAbs of high specificity can also determine the amount of phenotypic expression of polymorphic alleles and their metabolic role in drug and non-drug xenobiotic metabolism in heterozygote individuals. The inhibitory MAb might also identify allele-specific substrates of polymorphic human cytochrome P450s.
Laboratory of Molecular Carcinogenesis, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
K W
KW
Krausz
I
I
Goldfarb
T J
TJ
Yang
F J
FJ
Gonzalez
H V
HV
Gelboin
D000483:Alleles; D019943:Amino Acid Substitution / Q000235:genetics*; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000494:pharmacology*; D000918:Antibody Specificity; D001120:Arginine / Q000235:genetics / Q000378:metabolism; D001189:Aryl Hydrocarbon Hydroxylases; D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004008:Diclofenac / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D006825:Hybridomas; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D010616:Phenanthrenes / Q000378:metabolism; D011110:Polymorphism, Genetic / Q000235:genetics / Q000276:immunology; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism; D039181:Steroid 16-alpha-Hydroxylase; D013250:Steroid Hydroxylases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014044:Tolbutamide / Q000378:metabolism
D000911:Antibodies, Monoclonal; D065607:Cytochrome P-450 Enzyme Inhibitors; D007527:Isoenzymes; D010616:Phenanthrenes; D011994:Recombinant Proteins; D004008:Diclofenac; C031181:phenanthrene; D003577:Cytochrome P-450 Enzyme System; D001120:Arginine; D014044:Tolbutamide; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; D039181:Steroid 16-alpha-Hydroxylase; D003545:Cysteine