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Multiplex detection of food allergens and gluten

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An Erratum to this article was published on 26 July 2016

Abstract

To help safeguard the food supply and detect the presence of undeclared food allergens and gluten, most producers and regulatory agencies rely on commercial test kits. Most of these are ELISAs with a few being PCR-based. These methods are very sensitive and analyte specific, requiring different assays to detect each of the different food allergens. Mass spectrometry offers an alternative approach whereby multiple allergens may be detected simultaneously. However, mass spectrometry requires expensive equipment, highly trained analysts, and several years before a quantitative approach can be achieved. Using multianalyte profiling (xMAP®) technology, a commercial multiplex test kit based on the use of established antibodies was developed for the simultaneous detection of up to 14 different food allergens plus gluten. The assay simultaneously detects crustacean seafood, egg, gluten, milk, peanut, soy, and nine tree nuts (almond, Brazil nut, cashew, coconut, hazelnut, macadamia, pine nut, pistachio, and walnut). By simultaneously performing multiple tests (typically two) for each analyte, this magnetic bead-based assay offers built-in confirmatory analyses without the need for additional resources. Twenty-five of the assays were performed on buffer extracted samples, while five were conducted on samples extracted using reduced-denatured conditions. Thus, complete analysis for all 14 allergens and gluten requires only two wells of a 96-well microtiter plate. This makes it possible to include in a single analytical run up to 48 samples. All 30 bead sets in this multiplex assay detected 5 ng/mL of food allergen and gluten with responses greater than background. In addition, 26 of the bead sets displayed signal/noise ratios of five or greater. The bead-based design makes this 30-plex assay expandable to incorporate new antibodies and capture/detector methodologies by ascribing these new detectors to any of the unassigned bead sets that are commercially available.

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Acknowledgments

Our deepest gratitude is expressed to Mansour Samadpour, Ph.D. (IEH Laboratories & Consulting Group), Masahiro Shoji, Ph.D. (Morinaga Institute of Biological Sciences, Inc.), and Thomas Grace (Elution Technologies) for their openness and willingness to make resources available without which the development of this Multiplex Assay would not be possible. We would also like to acknowledge Ed Filardo, Ph.D. and Yoka Thomas who greatly assisted in the design and development of the method at Radix® BioSolutions. Appreciation is also expressed (posthumously) to Jesse Perry (FDA) for preparing the Black Tiger Prawn lyophilized powder used in this study. Thanks are also expressed to Lynn L. B. Rust, Ph.D. (NIH) and George C. Ziobro, Ph.D. (FDA) for interesting discussions.

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Authors and Affiliations

  1. Office of Regulatory Science, Center for Food Safety and Applied Nutrition (CFSAN), Food and Drug Administration, College Park, MD, 20740, USA

    Chung Y. Cho & Eric A.E. Garber

  2. Radix® BioSolutions, Georgetown, TX, 78626, USA

    William Nowatzke & Kerry Oliver

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  1. Chung Y. Cho
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  2. William Nowatzke
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  3. Kerry Oliver
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  4. Eric A.E. Garber
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Correspondence to Eric A.E. Garber.

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Cho, C.Y., Nowatzke, W., Oliver, K. et al. Multiplex detection of food allergens and gluten. Anal Bioanal Chem 407, 4195–4206 (2015). https://doi.org/10.1007/s00216-015-8645-y

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  • DOI: https://doi.org/10.1007/s00216-015-8645-y

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