J. Biochem, 1993, Vol. 114, No. 2 273-278
© 1993 Japanese Biochemical Society
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Preparation of Mouse Monoclonal Antibodies to Okadaic Acid and Their Binding Activity in Organic Solvents
*Division of Biochemistry, Iatron Laboratories Inc. 14600-6 Mitodai, Mito, Takomachi, Chiba 289-22
**Osaka Prefectural Institute of Public Health Higashi-ku, Osaka, Osaka 537
1To whom correspondence should be addressed.
Seven of 20 mouse monoclonal antibodies to OA, OA8–2, OA10–8, OA22–22, OA227–11, OA296–1, OA423–3, and OA958–2, were studied as to their binding to OA in organic solvents. OA423–3 (IgG2a-x) and OA958–2 (IgG1-x) in 90–100% methanol retained their binding activities with both immobilized and free antibodies. Whereas OA8–2 (IgG2a-x), OA10–8 (IgG1-x), OA22–22 (IgG2a-x), OA227–11 (IgG-x), and OA296–1 (IgM-x) did not bind to OA in over 50–60% methanol. The results of a non-competitive inhibition assay for OA indicated that in a methanolic or ethanolic solution, the binding ability of immobilized OA423–3 decreased as the concentration of each alcohol increased. The concentration of OA at the midpoint between the upper and lower plateaus of the inhibition curve was 0.18 ng/ml in 0% methanol and 570 ng/ml in 100%, respectively. In 0–50% of each of acetone, diethyl ether, and benzene in methanol, the binding ability of OA423–3 remained at the level in 100% methanol. OA958–2 showed similar binding properties to OA423–3. No relationship between the subclass of the immunoglobulin and the binding activity of the antibody in organic solvents was observed. These results indicate that the OA423–3 and OA958–2 antibodies are useful for the development of a new ELJSA method for OA in organic solvents.