Detection of hidden hazelnut protein in food by IgY-based indirect competitive enzyme-immunoassay

S. Baumgartner, M.G.E.G. Bremer, A.E.M. Kemmers - Voncken, N.G.E. Smits, W. Haasnoot, J. Banks, P. Reece, C. Danks, V. Tomkies, U. Immer, K. Schmitt, R. Krska

    Research output: Contribution to journalArticleAcademicpeer-review

    21 Citations (Scopus)

    Abstract

    The development of an indirect competitive enzyme-immunoassay for the detection of hidden hazelnut protein in complex food matrices is described. A sensitive and selective polyclonal antibody was raised by immunisation of laying hens with protein extracts from roasted hazelnuts. In contrast to traditional antibody generation in mammals, the antibody was not isolated from the blood of immunised mammals but from the egg yolk of immunised chickens. A standard calibration curve was optimised using immunoaffinity purified antibody extract and a coating antigen concentration of 10 ¿g ml¿1. One percent skim milk powder was chosen for blocking. The assay has a minimum detection limit of 10 ¿g l¿1, with an IC50 of 618 ¿g l¿1 when a 50 mM phosphate buffer at pH 7.5 and 10 mM sodium chloride is used as assay buffer. The cross reactivity testing shows a high specificity for hazelnut proteins and various foods and food additives were found to be non reactive except beans, sunflower seed or poppy seed.
    Original languageEnglish
    Pages (from-to)223-228
    JournalAnalytica Chimica Acta
    Volume520
    Issue number1/2
    DOIs
    Publication statusPublished - 2004

    Keywords

    • affinity-chromatography
    • anaphylaxis
    • antibodies
    • selection

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