Allergen Targets

Milk

Soy

Wheat

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Food Matrix

Cookies

Simultaneous detection of multi-allergens in an incurred food matrix using ELISA, multiplex flow cytometry and liquid chromatography mass spectrometry (LC-MS).

Gomaa, A., Boye, J.

Food chemistry (2015), 175, 585--92 DOI: 10.1016/j.foodchem.2014.12.017

Abstract

Food allergy is a public health concern and an important food safety issue. Food allergies affect up to 6% of infants and children and 4% of adults. The objective of this work was to determine differences in the detection of single and multiple allergens (i.e., casein, soy protein, and gluten) in an incurred food matrix before and after baking. Cookies were used as a model food system. Three methods, namely, multiplex assay (a new optimized method based on flow cytometry for multiple allergen analysis), enzyme-linked immunosorbent assay (ELISA) using commercial kits and LC-MS were used to detect allergens in the samples before and after baking. The ELISA kits performed well in detecting allergens in the raw samples with recoveries of 91-108%, 88-127% and 85-108% for casein, soy protein and gluten, respectively. Recoveries were poor for the baked cookies (67-90%, 66-95% and 66-88% for casein, soy protein and gluten, respectively). The multiplex flow cytometry assay permitted multiple allergen detection in the raw samples, with the following recoveries based on soluble protein: casein, 95-107%; soy protein, 92-97%, and gluten, 96-99%. Data for the baked cookies were as follows: casein, 84-90%; soy protein, 80-88%, and gluten, 80-90%. The LC-MS technique detected marker peptides that could be used to identify allergens in the baked food samples up to concentrations of 10 ppm for casein and soy protein, and 100 ppm for gluten. To the best of our knowledge, the current study is the first to compare ELISA, LC-MS and multiplex flow cytometry methods for the detection of multiple allergens simultaneously incurred in a model food system.

Reference raw data

Reference ID	Food	Protein	Peptide
28	Soy	Gly m 5	AIVILVINEGDANIELVGLK
28	Milk	Bos d 10	ALNEINQFYQK
28	Soy	Gly m 5	DLDIFLSIVDMNEGALLLPHFNSK
28	Soy	Gly m 6	HNIGQTSSPDIYNPQAGSVTTATSLDFPALSWLR
28	Milk	Bos d 9	HQGLPQEVLNENLLR
28	Soy	Gly m 6	HQQEEENEGGSILSGFTLEFLEHAFSVDK
28	Milk	Bos d 10	LNFLK
28	Milk	Bos d 10	NAVPITPTLNR
28	Soy	Gly m 6	QNIGQNSSPDIYNPQAGSITTATSLDFPALWLLK
28	Soy	Gly m 6	TNDRPSIGNLAGANSLLNALPEEVIQHTFNLK
28	Soy	Gly m 6	TNDTPMIGTLAGANSLLNALPEEVIQHTFNLK
28	Soy	Gly m 6	VLIVPQNFVVAAR
28	Milk	Bos d 12	YIPIQYVLSR