Hapten Design and Antibody Preparation on Immunoassay toward the Insecticide Oxamyl
Shiro MIYAKE, Kosuke MORIMUNE, Yuki YAMAGUCHI, Katsuya OHDE, Mitsuyasu KAWATA, Shunichi TAKEWAKI and Yohjiro YUASA
J. Pestic. Sci.
25, 10-17 (2000)
To detect small amount of oxamyl, indirect competitive enzyme-linked immunosorbent assays (indirect C-ELISAs) were established. Four haptens for immunogen and three haptens for coating antigen on indirect C-ELISA were synthesized and their polyclonal (PoAbs) and monoclonal antibodies (MoAbs) were prepared. Hapten N, N-dimethyl-2-(5-carboxypentyl)carbamoyloxyimino-2-(methylthio)acetamide was the most effective to raise antibodies toward oxamyl out of haptens for immunogen. MoAbs prepared with this hapten were more highly reactive than PoAbs. In particular, MoAb OXM 6กพ8 showed the highest reactivity. When N,N-dimethyl-2-(4-carboxycyclohexyl)carbamoyloxyimino-2-(methylthio)acetamide was used for coating antigen on indirect C-ELISA with OXM 6กพ8, the highest sensitivity was observed and the concentration for 50% inhibition value showed 3.6ng/ml. Methanol extract of potato showed no effect on the reactivity.
