ANTIBODIES TO A SYNTHETIC PEPTIDE THAT REACT WITH FLAVIN-CONTAINING MONOOXYGENASE (HLFMO3) IN HUMAN HEPATIC MICROSOMES

Flavin-containing monooxygenases (FMOs) catalyze the oxidation of a di verse array of xenobiotic compounds. The purpose of this investigation was to develop a specific immunological probe to human hepatic flavin -containing monooxygenase (HLFMO3). An oligopeptide corresponding to a mino acid residues 257-270 of HLFMO3 was coupled to keyhole limpet hem ocyanin (KLH) through the sulfhydryl group of a cysteine residue added to the amino-terminus of the peptide. This peptide-KLH conjugate was used to generate a polyclonal antibody. The resulting immunoglobulin s howed specific Western blot reactivity with HLFMO3 protein in human he patic microsomes, the same protein that is recognized by a polyclonal antibody directed against macaque liver FMO. These findings demonstrat e that an antibody directed against a synthetic peptide derived from H LFMO3 can be easily produced in large quantities and used in studies f or the immunodetection and immunoquantification of HLFMO3. This is als o the first antipeptide antibody directed against an FMO of any specie s. (C) 1997 Elsevier Science Inc.