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Development of a competitive liposome-based lateral flow assay for the rapid detection of the allergenic peanut protein Ara h1

Wen, Hsiao-Wei ; Borejsza-Wysocki, Wlodzimierz ; DeCory, Thomas ; Durst, Richard

In: Analytical and Bioanalytical Chemistry, 2005, vol. 382, no. 5, p. 1217-1226

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    Titre
    • Development of a competitive liposome-based lateral flow assay for the rapid detection of the allergenic peanut protein Ara h1
    Auteur
    • Wen, Hsiao-Wei. Department of Food Science and Technology, Cornell University, 630 West North Street, NY 14456-0462, Geneva, USA
    • Borejsza-Wysocki, Wlodzimierz. Department of Food Science and Technology, Cornell University, 630 West North Street, NY 14456-0462, Geneva, USA
    • DeCory, Thomas. Department of Food Science and Technology, Cornell University, 630 West North Street, NY 14456-0462, Geneva, USA
    • Durst, Richard. Department of Food Science and Technology, Cornell University, 630 West North Street, NY 14456-0462, Geneva, USA
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Analytical and Bioanalytical Chemistry, 2005, vol. 382, no. 5, p. 1217-1226. Springer-Verlag; http://www.springer.de
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:312447
    Summary
    A competitive lateral flow assay for detecting the major peanut allergen, Ara h1, has been developed. The detector reagents are Ara h1-tagged liposomes, and the capture reagents are anti-Ara h1 polyclonal antibodies. Two types of rabbit polyclonal antibodies were raised either against the entire Ara h1 molecules (anti-Ara h1 Ab) or against an immunodominant epitope on Ara h1 (anti-peptide Ab). All of them reacted specifically with Ara h1 in Western Blot against crude peanut proteins. Moreover, the anti-Ara h1 Ab was chosen for this assay development because of its highest immunoactivity to Ara h1-tagged liposomes in the lateral flow assay. The calculated limit of detection (LOD) of this assay is 0.45μgmL−1 of Ara h1 with a dynamic range between 0.1 and 10μgmL−1 of Ara h1 in buffer. Additionally, the visually determined detection range is from 1 to 10μgmL−1 of Ara h1 in buffer. Results using this assay can be obtained within 30min without the need of sophisticated equipment or techniques; therefore, this lateral flow assay has the potential to be a cost-effective, fast, simple, and sensitive method for on-site screening of peanut allergens