Monoclonal antibody-directed immunopurification and identification of cytochromes P-450.
Several rat liver cytochromes P-450 have been substantially purified in a one-step immunoadsorption procedure using Sepharose-bound monoclonal antibodies (MAbs) to the major forms of rat liver cytochrome P-450 induced by 3-methylcholanthrene and phenobarbital (MC-P-450 and PB-P-450, respectively). When mixed with solubilized rat liver microsomes the immunoadsorbent based on the MAb to MC-P-450 binds two polypeptides of MW 56,000 and 57,000 while the immunoadsorbent made with the MAb to PB-P-450 absorbs a species of MW 54,000. These polypeptides are readily desorbed by 0.1 M glycine (pH 3.0). Isolation of MAb-specific cytochrome P-450 isozymes by this method has applications in numerous phases of cytochrome P-450 research.
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D004790:Enzyme Induction; D006825:Hybridomas / Q000276:immunology; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D051379:Mice; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism*; D010634:Phenobarbital / Q000494:pharmacology; D010954:Plasmacytoma / Q000276:immunology; D051381:Rats; D011919:Rats, Inbred Strains
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D008748:Methylcholanthrene; D003577:Cytochrome P-450 Enzyme System; D010634:Phenobarbital