Quantitative assessment of factors affecting the sensitivity of a competitive immunomicroarray for pesticide detection

Erik Belleville, Hans Martin Dufva, J., Aamund, L. Bruun, Claus Bo Vöge Christensen

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    Analytical protein microarrays offering highly parallel analysis can become an invaluable tool for a wide range of immunodiagnostic applications. Here we describe factors that influence the sensitivity of a competitive immunomicroarray that quantifies small molecules; in this case, the pesticides dichlobenil metabolite 2,6-dichlorobenzamide (BAM) and atrazine. Free pesticide concentrations in solution are quantified by the competitive binding of fluorescence-conjugated monoclonal antibodies to either surface-immobilized pesticide hapten-protein conjugates or pesticides in solution. We investigated the influence of antibody labeling techniques, microarray substrates, and spotting and incubation buffers. The results showed that microarrays immobilized on EasySpot or in-house fabricated agarose substrates printed with Genetix Amine Spotting Solution resulted in optimum results when the arrays were incubated with the sample/antibodies diluted in a Tris buffer supplemented with 0.05% each bovine serum albumin (BSA) and Tween(R) 20. Furthermore, the application of directly labeled primary antibodies allowed for better sensitivity compared to secondary polyclonal antibody quantification.
    Original languageEnglish
    JournalBioTechniques
    Volume35
    Issue number5
    Pages (from-to)1044
    ISSN0736-6205
    Publication statusPublished - 2003

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