Abstract
A specific antibody has been produced for developing immunoaffinity chromatography of optically active bufuralol and its 1′-oxidized metabolites. Spleen cells from a BALB/c mouse, which had been immunized with the (1R)- or (1S)-1′-oxobufuralol O-carboxymethyl oxime-bovine serum albumin conjugate, were fused with P3/NS1/1-Ag4-1 myeloma cells. After screening by enzyme-linked immunosorbent assay (ELISA) using a β-galactosidase-labeled antigen, one for (1R)-bufuralol and three kinds for (1S)-bufuralol of antibody-secreting hybridoma clones were established. The monoclonal antibodies, Ab-R-12 and Ab-S-1,were determined to have favorable characteristics for preparing the immunosorbent, because each of them was specific for (1R)- or (1S)-bufuralols and its corresponding 1′-oxidized metabolites but showed significantly low cross-reactivities with their antipodes including 1′-oxidized metabolites.
