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

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Abstract

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 0.45 μg mL−1 of Ara h1 with a dynamic range between 0.1 and 10 μg mL−1 of Ara h1 in buffer. Additionally, the visually determined detection range is from 1 to 10 μg mL−1 of Ara h1 in buffer. Results using this assay can be obtained within 30 min 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.

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Acknowledgements

The authors thank Dr. Antje Baeumner and Dr. Randy Worobo of Cornell University for their helpful suggestions on this research, and for the Vitasoy & Lo Fellowship awarded to Hsio-Wei Wen.

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Authors and Affiliations

  1. Department of Food Science and Technology, Cornell University, 630 West North Street, Geneva, NY 14456-0462, USA

    Hsiao-Wei Wen, Wlodzimierz Borejsza-Wysocki, Thomas R. DeCory & Richard A. Durst

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  1. Hsiao-Wei Wen
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  2. Wlodzimierz Borejsza-Wysocki
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  3. Thomas R. DeCory
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  4. Richard A. Durst
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Correspondence to Richard A. Durst.

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Wen, HW., Borejsza-Wysocki, W., DeCory, T.R. et al. Development of a competitive liposome-based lateral flow assay for the rapid detection of the allergenic peanut protein Ara h1. Anal Bioanal Chem 382, 1217–1226 (2005). https://doi.org/10.1007/s00216-005-3292-3

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  • DOI: https://doi.org/10.1007/s00216-005-3292-3

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