A flow-cytometry-based method for detecting simultaneously five allergens in a complex food matrix

• Published in: Journal of food science and technology 2016-12, Vol.53 (12), p.4179-4186
• Main Authors: Otto, Gaetan, Lamote, Amandine, Deckers, Elise, Dumont, Valery, Delahaut, Philippe, Scippo, Marie-Louise, Pleck, Jessica, Hillairet, Caroline, Gillard, Nathalie
• Format: Article
• Language: English
• Subjects: Allergens; Allergies; Analysis; Antibodies; Arachis hypogaea; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Contaminants; cookies; Crustacea; Crustaceans; eggs; enzyme-linked immunosorbent assay; Flow cytometry; Food; Food allergies; food industry; Food matrix; Food processing industry; Food quality; Food Science; Gluten; Immunology; Ingredients; inhibitory concentration 50; issues and policy; Life sciences; Methods; microbeads; milk; Nutrition; Original; Original Article; peanuts; Pollution control; Product recalls; Proteins; Quality control; Sciences des denrées alimentaires; Sciences du vivant; sorbents; Studies
• ISSN: 0022-1155; 0975-8402; 0975-8402
• DOI: 10.1007/s13197-016-2402-x

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.