Development of an ELISA for the organophosphorus insecticide chlorpyrifos

A selective enzyme-linked immunosorbent assay (ELISA) for the insecticide chlorpyrifos was developed. Four haptens for chlorpyrifos were synthesized and two of them were used as immunogens after coupling to keyhole limpet hemocyanin by two different approaches. Rabbits were immunized with either of them and the sera were screened against 4 haptens coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigen-coated ELISA was developed, which shows an I-50 of I-60 ppb with a detection limit of 10 ppb. An antibody-coated ELISA was also developed, which shows an I-50 of 20 ppb with a detection limit of 0.1 ppb. The antibodies showed negligible cross-reactivity with other organophosphorus pesticides except for insecticides chlorpyrifos-methyl and bromophos-ethyl, which makes these assays suitable for the selective detection of chlorpyrifos.