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Development of an Enzyme-Linked Immunosorbent Assay for the Organophosphorus Fungicide Tolclofos-methyl

  • Park, Kyung-Yi (Department of Biochemistry and Institute of Natural Sciences, Yeungnam University) ;
  • Park, Won-Chul (Department of Biochemistry and Institute of Natural Sciences, Yeungnam University) ;
  • Kim, Yoo-Jung (Department of Biochemistry and Institute of Natural Sciences, Yeungnam University) ;
  • Lee, Yong-Tae (Department of Biochemistry and Institute of Natural Sciences, Yeungnam University)
  • Published : 2003.03.20

Abstract

A simple synthetic method for haptens of organophosphorus (OP) pesticides with a spacer arm (aminocarboxylic acid) attached at the pesticide thiophosphate group was developed and was applied to the synthesis of haptens for the OP fungicide tolclofos-methyl. Using the haptens, a selective enzyme-linked immunosorbent assay (ELISA) for tolclofos-methyl was developed. One of the haptens was coupled to BSA to use as an immunogen. Rabbits were immunized with this conjugate to obtain polyclonal antibodies to tolclofos-methyl. The antisera were screened against another hapten coupled to ovalbumin (OVA). Using the serum with highest specificity, an antigen-coated ELISA was developed, which showed an $IC_{50}$ of 160 ng/mL with the detection limit of 20 ng/mL. The antibodies showed negligible cross-reactivity with other OP pesticides. An antibody-coated ELISA was also developed, which showed an $IC_{50}$ of 410 ng/mL with a detection limit of 130 ng/mL.

Keywords

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