title: Quantitative double antibody sandwich ELISA for the determination of gliadin.
abstract: A sensitive double antibody sandwich ELISA (DAS-ELISA) based on chicken anti-gliadin IgY and biotinylated monoclonal antibody (mAb) was developed for the quantification of gliadin in foods. The anti-gliadin IgY and mAb specifically detected gliadin in wheat, barley, and rye by indirect ELISA and Western-blot assay. Using anti-gliadin IgY as capture antibody and biotinylated mAb as detecting antibody, the sensitivity of DAS-ELISA has a linear standard range of 4-40 ng/mL, showing that the limit of detection (LOD) corresponds to 4 ng/mL gliadin in assay buffer, equivalent to 0.8 ppm in foods. The intra-assay expressed as percentage of coefficients of variation (%CV) was 7.25% average of six food samples. The interassay precision was 9.51% in food samples. The combination of anti-gliadin IgY and biotinylated mAb in the DAS-ELISA provides a reliable, sensitive, and inexpensive tool for the detection of gliadin in gluten-free and gluten-containing food products.
authors.affiliation: Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada.
authors.forename: Naiyana
authors.initials: N
authors.lastname: Gujral
authors.forename: Mavanur R
authors.initials: MR
authors.lastname: Suresh
authors.forename: Hoon H
authors.initials: HH
authors.lastname: Sunwoo
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005903:Gliadin / Q000032:analysis* / Q000276:immunology; D007136:Immunoglobulins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; C016512:IgY; D007136:Immunoglobulins; D005903:Gliadin