Abstract
To avoid carry-over contamination with allergens, food manufacturers implement quality control strategies relying primarily on detection of allergenic proteins by ELISA. Although sensitive and specific, this method allowed detection of only one allergen per analysis and effective control policies were thus based on multiplying the number of tests done in order to cover the whole range of allergens. We present in this work an immunoassay for the simultaneous detection of milk, egg, peanut, mustard and crustaceans in cookies samples. The method was based on a combination of flow cytometry with competitive ELISA where microbeads were used as sorbent surface. The test was able to detect the presence of the five allergens with median inhibitory concentrations (IC50) ranging from 2.5 to 15 mg/kg according to the allergen to be detected. The lowest concentrations of contaminants inducing a significant difference of signal between non-contaminated controls and test samples were 2 mg/kg of peanut, 5 mg/kg of crustaceans, 5 mg/kg of milk, 5 mg/kg of mustard and 10 mg/kg of egg. Assay sensitivity was influenced by the concentration of primary antibodies added to the sample extract for the competition and by the concentration of allergenic proteins bound to the surface of the microbeads.
Similar content being viewed by others
References
Alvarez PA, Boye JI (2012) Food production and processing considerations of allergenic food ingredients: a review. J Allergy (Cairo) 2012:746125. doi:10.1155/2012/746125
Bienenmann-Ploum ME, Huet AC, Campbell K, Fodey TL, Vincent U, Haasnoot W, Delahaut P, Elliott CT, Nielen MW (2012) Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six coccidiostats in feed and eggs. Anal Bioanal Chem 404(5):1361–1373
Crevel RW, Kerkhoff MA, Koning MM (2000) Allergenicity of refined vegetable oils. Food Chem Toxicol 38(4):385–393
Figueroa J, Blanco C, Dumpiérrez AG, Almeida L, Ortega N, Castillo R, Navarro L, Pérez E, Gallego MD, Carrillo T (2005) Mustard allergy confirmed by double-blind placebo-controlled food challenges: clinical features and cross-reactivity with mugwort pollen and plant-derived foods. Allergy 60(1):48–55
Gendel SM (2013) The regulatory challenge of food allergens. J Agric Food Chem 61(24):5634–5637
Gomaa A, Boye J (2015) Simultaneous detection of multi-allergens in an incurred food matrix using ELISA, multiplex flow cytometry and liquid chromatography mass spectrometry (LC-MS). Food Chem 15(175):585–592
Gomaa A, Ribereau S, Boye J (2012) Detection of allergens in a multiple allergen matrix and study of the impact of thermal processing. J Nutr Food Sci S9:001
Gupta R, Sheikh A, Strachan DP, Anderson HR (2007) Time trends in allergic disorders in the UK. Thorax 62(1):91–96
Haasnoot W, du Pré JG (2007) Luminex-based triplex immunoassay for the simultaneous detection of soy, pea, and soluble wheat proteins in milk powder. J Agric Food Chem 55(10):3771–3777
Hermanson G (2008) Bioconjugate techniques, 2nd edn. Academic Press, Cambridge, pp 802–805
Huet AC, Charlier C, Singh G, Godefroy SB, Leivo J, Vehniäinen M, Nielen MW, Weigel S, Delahaut P (2008) Development of an optical surface plasmon resonance biosensor assay for (fluoro) quinolones in egg, fish, and poultry meat. Anal Chim Acta 623(2):195–203
Kirsch S, Fourdrilis S, Dobson R, Scippo ML, Maghuin-Rogister G, De Pauw E (2009) Quantitative methods for food allergens: a review. Anal Bioanal Chem 395:57–67
Koppelman SJ, Vlooswijk R, Bottger G, Van Duijn G, Van der Schaft P, Dekker J, Van Bergen H (2007) Development of an enzyme-linked immunosorbent assay method to detect mustard protein in mustard seed oil. J Food Prot 70(1):179–183
Monaci L, Visconti A (2010) Immunochemical and DNA-based methods in food allergen analysis and quality assurance perspectives. Trends Food Sci Technol 21(6):272–283
Peters J, Bienenmann-Ploum M, de Rijk T, Haasnoot W (2011) Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed. Mycotoxin Res 27(1):63–72
Peters J, Thomas D, Boers E, de Rijk T, Berthiller F, Haasnoot W, Nielen MW (2013) Colour-encoded paramagnetic microbead-based direct inhibition triplex flow cytometric immunoassay for ochratoxin A, fumonisins and zearalenone in cereals and cereal-based feed. Anal Bioanal Chem 405(24):7783–7794
Poms RE, Klein CL, Anklam E (2004) Methods for allergen analysis in food: a review. Food Addit Contam 21(1):1–31
Taylor SL, Baumert JL, Kruizinga AG, Remington BC, Crevel RW, Brooke-Taylor S, Allen KJ, Allergen Bureau of Australia & New Zealand, Houben G (2014) Establishment of reference doses for residues of allergenic foods: report of the VITAL expert panel. Food Chem Toxicol 63:9–17
Verhoeckx KC, Vissers YM, Baumert JL, Faludi R, Feys M, Flanagan S, Herouet-Guicheney C, Holzhauser T, Shimojo R, van der Bolt N, Wichers H, Kimber I (2015) Food processing and allergenicity. Food Chem Toxicol 80:223–240
Worm M, Timmermans F, Moneret-Vautrin A, Muraro A, Yman IM, Lövik M, Hattersley S, Crevel R (2010) Towards a European registry of severe allergic reactions: current status of national registries and future needs. Allergy 65(6):671–680
Zitouni N, Errahali Y, Metche M, Kanny G, Moneret-Vautrin DA, Nicolas JP, Fremont S (2000) Influence of refining steps on trace allergenic protein content in sunflower oil. J Allergy Clin Immunol 106(5):962–967
Acknowledgements
This project was financially supported by a grant of the Walloon Region (Belgium): Collective Research Program–convention 1217554.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Otto, G., Lamote, A., Deckers, E. et al. A flow-cytometry-based method for detecting simultaneously five allergens in a complex food matrix. J Food Sci Technol 53, 4179–4186 (2016). https://doi.org/10.1007/s13197-016-2402-x
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13197-016-2402-x