Analytical Sciences


Abstract − Analytical Sciences, 16(1), 31 (2000).

13Production and Characterization of Monoclonal Antibodies Recognizing the Asymmetric Center of Optically Active Bufuralol and Its Metabolites
Shigeo IKEGAWA, Ni Made Ria ISRIYANTHI, Norihiro KOBAYASHI, Shinichi MIYAIRI, and Junichi GOTO *
Graduate School of Pharmaceutical Sciences, Tohoku University, Aobayama, Sendai, 980-8578, Japan
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 beta-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.