Development of an enzyme-linked immunosorbent assay for carbaryl

Maria Pilar Marco, Shirley J. Gee, Hong M. Cheng, Zeng Y. Liang, Bruce D. Hammock

Research output: Contribution to journalArticle

70 Scopus citations

Abstract

Seven rabbit antisera were obtained by immunization with either N-(1-naphthylacetyl)-6-aminohexanoic acid (3) or 1-(1-naphthyl)-3-(5-carboxypentyl)urea (8) conjugated to keyhole limpet hemocyanin (KLH). From these sera Ab2114 (against 8KLH) was used for the optimization of an enzyme-linked immunosorbent assay (ELISA) for the determination of the insecticide carbaryl. An I50 of 2-5 ng/mL and a detection limit of 0.2 ng/mL were obtained using N-(2-naphthoyl)-6-aminohexanoic acid (5) coupled to conalbumin as a coating antigen. No interference with naphthol, the main degradation product of carbaryl, was observed. An enzyme tracer was prepared by covalently linking hapten 5 with alkaline phosphatase. When the ELISA was performed using a format involving coating the plate with antibodies, an I50 of 0.4-0.6 ng/mL and a detection limit of 0.05 ng/mL were obtained. Preliminary studies with different samples showed that this immunoassay can be used for the determination of carbaryl in water, soil, body fluids, and food samples. This paper demonstrates the flexibility of using stable derivatives of a target compound such as carbaryl to generate antibodies and a sensitive ELISA for molecules containing functionalities susceptible to chemical hydrolysis and biodegradation.

Original languageEnglish (US)
Pages (from-to)423-430
Number of pages8
JournalJournal of Agricultural and Food Chemistry
Volume41
Issue number3
StatePublished - 1993

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Food Science
  • Chemistry (miscellaneous)

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    Marco, M. P., Gee, S. J., Cheng, H. M., Liang, Z. Y., & Hammock, B. D. (1993). Development of an enzyme-linked immunosorbent assay for carbaryl. Journal of Agricultural and Food Chemistry, 41(3), 423-430.