Analytical Sciences

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.

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