Production and characterization of monoclonal antibodies recognizing the asymmetric center of optically active bufuralol and its metabolites

A specific antibody has been produced for developing immunoaffinity chromat ography of optically active bufuralol and its 1'-oxidized metabolites. Sple en cells from a BALB/c mouse, which had been immunized with the (1R)- or (1 S)-1'-oxobufuralol O-carboxymethyl oxime-bovine serum albumin conjugate, we re fused with P3/NS1/1-Ag4-1 myeloma cells. After screening by enzyme-linke d immunosorbent assay (ELISA) using a beta-galactosidase-labeled antigen, o ne for (1R)bufuralol and three kinds for (1S)-bufuralol of antibody-secreti ng hybridoma clones were established. The monoclonal antibodies, Ab-R-12 an d Ab-S-1,were determined to have favorable characteristics for preparing th e immunosorbent, because each of them was specific for (1R)- or (1S)-bufura lols and its corresponding 1'-oxidized metabolites but showed significantly low cross-reactivities with their antipodes including 1'-oxidized metaboli tes.