A simple and specific noncompetitive ELISA method for HT-2 toxin detection

Henri O. Arola, Antti Tullila, Alexis V. Nathanail, Tarja K. Nevanen

    Research output: Contribution to journalArticleScientificpeer-review

    14 Citations (Scopus)

    Abstract

    We developed an HT-2 toxin-specific simple ELISA format with a positive read-out. The assay is based on an anti-immune complex (IC) scFv antibody fragment, which is genetically fused with alkaline phosphatase (AP). The anti-IC antibody specifically recognizes the IC between a primary anti-HT-2 toxin Fab fragment and an HT-2 toxin molecule. In the IC ELISA format, the sample is added together with the scFv-AP antibody to the ELISA plate coated with the primary antibody. After 15 min of incubation and a washing step, the ELISA response is read. A competitive ELISA including only the primary antibody recognizes both HT-2 and T-2 toxins. The anti-IC antibody makes the assay specific for HT-2 toxin, and the IC ELISA is over 10 times more sensitive compared to the competitive assay. Three different naturally contaminated matrices: wheat, barley and oats, were used to evaluate the assay performance with real samples. The corresponding limits of detection were 0.3 ng/mL (13 µg/kg), 0.1 ng/mL (4 µg/kg) and 0.3 ng/mL (16 µg/kg), respectively. The IC ELISA can be used for screening HT-2 toxin specifically and in relevant concentration ranges from all three tested grain matrices.
    Original languageEnglish
    Article number145
    JournalToxins
    Volume9
    Issue number4
    DOIs
    Publication statusPublished - 2017
    MoE publication typeA1 Journal article-refereed

    Keywords

    • Alkaline phosphatase
    • Cereal grains
    • ELISA
    • Fab
    • Fusion protein
    • HT-2 toxin
    • Noncompetitive
    • Recombinant antibodies
    • scFv

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