Preparation of mouse monoclonal antibodies to okadaic acid and their binding activity in organic solvents.
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 (IgG1-kappa) and OA958-2 (IgG1-kappa) in 90-100% methanol retained their binding activities with both immobilized and free antibodies. Whereas OA8-2 (IgG2a-kappa), OA10-8 (IgG1-kappa), OA22-22 (IgG2a-kappa), OA227-11 (IgG1-kappa), and OA296-1 (IgM-kappa) 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 ELISA method for OA in organic solvents.
Matsuura
S
S
Division of Biochemistry, Iatron Laboratories Inc., Chiba.
Hamano
Y
Y
Kita
H
H
Takagaki
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D001665:Binding Sites; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000431:Ethanol / Q000737:chemistry; D004988:Ethers, Cyclic / Q000032:analysis / Q000276:immunology*; D006825:Hybridomas; D008387:Marine Toxins / Q000276:immunology; D000432:Methanol / Q000737:chemistry; D051379:Mice; D019319:Okadaic Acid; D012997:Solvents / Q000737:chemistry
D000911:Antibodies, Monoclonal; D004988:Ethers, Cyclic; D008387:Marine Toxins; D012997:Solvents; D019319:Okadaic Acid; D000431:Ethanol; D000432:Methanol