Hapten heterology for a specific and sensitive indirect enzyme-linked immunosorbent assay for organophosphorus insecticide fenthion

Abstract

Five haptens with different spacer-arm attachment sites on the structure of the organophosphorus insecticide fenthion were designed and synthesized. All of the haptens were conjugated with ovalbumin (OVA) for the coating antigen, and three haptens containing all or most of the structure of fenthion were conjugated with bovine serum albumin (BSA) for the immunogen. Six polyclonal antisera were raised against the three BSA conjugates, and 30 antibody/coating conjugate combinations were selected for studies of assay sensitivity and specificity for fenthion. The study revealed the best combination with high sensitivity ( I 50 of 0.08 ng mL −1) and high assay specificity, which indicated that when structural difference between the analyte and an immunizing hapten is less than that between a coating hapten and the immunizing hapten, a high sensitive enzyme-linked immunosorbent assay (ELISA) in the heterologous system may stand a good chance to be developed. The immunity results showed that heterology in the hapten spacer-arm attachment site of the immunogen could achieve a remarkable improvement in the quantity, sensitivity, and/or specificity of antibody, and that the moiety of an analyte, which is the same as the moiety near/on the immunizing spacer-arm hapten attachment site, contributes greatly to the interaction of antibody and hapten.