title: Implementation of an Enzyme Linked Immunosorbent Assay for the Quantification of Allergenic Egg Residues in Red Wines Using Commercially Available Antibodies.
abstract: Since the early 2000s, labeling of potentially allergenic food components to protect people who suffer from food allergies is compulsory in numerous industrialized countries. In Europe, milk and egg components used during the winemaking process must be indicated on the label since July 1, 2012. Several ELISA procedures have been developed to detect allergenic residues in wines. However, the complexity of the wine matrix can inhibit the immunoenzymatic reaction. The aim of this study was to implement an ELISA assay for the detection of ovalbumin in red wines using commercially available antibodies. The specificity of the acquired antibodies and the absence of cross reactivity were assessed by immunoblotting and ELISA. An ELISA assay with a LOD of 14.2 μg/L and a LOQ of 56.4 μg/L of ovalbumin in aqueous solution was obtained. Differences in ELISA signals were observed when analyzing various fining agents, although reproducible conformation of the antigen could be reached for the comparison of ovalbumin and Ovicolle. The differences between samples in terms of pH could be leveled but the inhibition of the ELISA signal, positively correlated to the tannin content of the wines, could not be suppressed. Thus, standard curves of ovalbumin in several wines were obtained by relative quantification. The control steps and the difficulties encountered presented in this study should be considered by anybody working toward the development of ELISA assays for the detection of allergenic residues in complex food matrices.
authors.affiliation: Inst. of Plant Production Sciences, Swiss Federal Research Station Agroscope, Route de Duillier 50, Case Postale 1012, 1260, NYON, Switzerland.
authors.forename: Carole
authors.initials: C
authors.lastname: Koestel
authors.affiliation: Inst. of Plant Production Sciences, Swiss Federal Research Station Agroscope, Route de Duillier 50, Case Postale 1012, 1260, NYON, Switzerland.
authors.forename: Céline
authors.initials: C
authors.lastname: Simonin
authors.affiliation: Inst. of Food Sciences, Swiss Federal Research Station Agroscope, Route de Duillier 50, Case Postale 1012, 1260, NYON, Switzerland.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Belcher
authors.affiliation: Inst. of Plant Production Sciences, Swiss Federal Research Station Agroscope, Route de Duillier 50, Case Postale 1012, 1260, NYON, Switzerland.
authors.forename: Johannes
authors.initials: J
authors.lastname: Rösti
mesh_terms: D000485:Allergens / Q000032:analysis*; D000818:Animals; D000906:Antibodies; D003429:Cross Reactions; D004531:Eggs; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005060:Europe; D005506:Food Contamination / Q000032:analysis; D005512:Food Hypersensitivity / Q000276:immunology; D006801:Humans; D008892:Milk / Q000737:chemistry; D010047:Ovalbumin / Q000032:analysis* / Q000276:immunology; D013634:Tannins / Q000032:analysis; D014920:Wine / Q000032:analysis*
chemical_list: D000485:Allergens; D000906:Antibodies; D013634:Tannins; D010047:Ovalbumin
keywords: ELISA; allergen; egg; method validation; wine