Influence of the ELISA format and the hapten-enzyme conjugate on the sensitivity of an immunoassay for s-triazine herbicides using monoclonal antibodies

Peter Schneider, Bruce D. Hammock

Research output: Contribution to journalArticle

135 Scopus citations

Abstract

A variety of formats for enzyme-linked immunosorbent assays have been examined for the analysis of environmental samples using a single monoclonal antibody raised to the herbicide atrazine. The most favorable combination of several ELISA formats and a variety of synthesized hapten-enzyme conjugates resulted in a highly sensitive enzyme immunoassay with 50 % inhibition (I50) at 0.25 μg/L and a detection limit of 0.03 μg/L. This assay is the most sensitive atrazine assay, based on monoclonal antibodies published so far, and can be performed in less than 1 h. All formats were based on the use of hapten-enzyme conjugates (enzyme tracers). Direct coating and precoating with goat anti-mouse antibodies or protein A were used. The formats using precoating steps turned out to be the most sensitive. Enzyme tracers based on two different atrazine and simazine haptens each with two different spacers (C3 or C6) were conjugated using either alkaline phosphatase or horseradish peroxidase. The atrazine derivative with the C6 spacer arm conjugated to horseradish peroxidase provided highest sensitivity.

Original languageEnglish (US)
Pages (from-to)525-530
Number of pages6
JournalJournal of Agricultural and Food Chemistry
Volume40
Issue number3
StatePublished - 1992

ASJC Scopus subject areas

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

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