Monoclonal antibody-based enzyme-linked immunosorbent assays for the organophosphorus insecticide O-ethyl O-4-nitrophenyl phenylphosphonothioate (epn)

Jee Youn Shim, Young Ah Kim, Yong Tae Lee, Bruce D. Hammock, Hye Sung Lee

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

This study aimed at developing competitive direct and indirect enzyme-linked immunosorbent assays (ELISAs) for the organophosphorus insecticide O-ethyl O-4-nitrophenyl phenylphosphonothioate (EPN) using a monoclonal antibody (mAb). Of the five EPN derivatives (haptens) prepared for use as an immunogen or as a competitor, two of them were used as the immunogen for the production of the mAbs. By using the antibody with the highest specificity and a coating antigen (hapten-OVA conjugate), a competitive indirect ELISA was developed, which showed an IC50 of 2.9 ng/mL with a detection limit of 0.3 ng/mL. A competitive direct ELISA using a different antibody and an enzyme tracer was also developed, which showed an IC50 of 0.6 ng/mL with a detection limit of 0.09 ng/mL. The mAbs in both assays showed negligible cross-reactivity with other organophosphorus pesticides. The recoveries of EPN from spiked samples determined by the developed ELISA ranged from 59 to 143%. Dilution of the samples improved the recovery. The assay performance of the present ELISAs based on the mAb was compared with that of the EPN ELISAs based on polyclonal antibodies (pAbs) that had been developed previously and was found to be better in dynamic response.

Original languageEnglish (US)
Pages (from-to)5241-5247
Number of pages7
JournalJournal of Agricultural and Food Chemistry
Volume58
Issue number9
DOIs
StatePublished - May 12 2010

Keywords

  • ELISA
  • EPN
  • Immunoassay
  • Insecticide
  • Monoclonal antibody

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Chemistry(all)

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