title: Development of sandwich ELISA for testing bovine β-lactoglobulin allergenic residues by specific polyclonal antibody against human IgE binding epitopes.
abstract: Bovine β-lactoglobulin (BLG) is the main allergen in cows' milk, and the most commonly used method for detecting BLG is enzyme-linked immunosorbent assay (ELISA). However, antibodies used in commercial ELISA kits do not recognize specifically BLG IgE epitopes. Here, an antibody specific to IgE linear epitopes for BLG was used to develop a sandwich ELISA using a rabbit anti-BLG polyclonal antibody. The linear range for BLG detection was 31.25-8000ng/mL and limit of detection was 1.96ng/mL. BLG content in dairy samples was determined, and there was a good agreement between this immunoassay and reversed-phase high-performance liquid chromatography with high recovery. Additionally, BLG content in food samples had an average recovery of 104.25%. Allergenic residues were also detected in hydrolyzed infant formulas. The method developed could be a practical approach to determine BLG and its allergenic residues in food with a high degree of sensitivity, reliability and recovery.
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China; School of Food Science & Technology, Nanchang University, Nanchang 330031, PR China; Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, PR China.
authors.forename: Shengfa
authors.initials: S
authors.lastname: He
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China; School of Food Science & Technology, Nanchang University, Nanchang 330031, PR China.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China; School of Food Science & Technology, Nanchang University, Nanchang 330031, PR China.
authors.forename: Jinyan
authors.initials: J
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China.
authors.forename: Ping
authors.initials: P
authors.lastname: Tong
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China; Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, PR China. Electronic address: chenhongbing@ncu.edu.cn.
authors.forename: Hongbing
authors.initials: H
authors.lastname: Chen
mesh_terms: D000485:Allergens / Q000032:analysis / Q000276:immunology; D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D005260:Female; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis* / Q000276:immunology; D041943:Infant Formula / Q000032:analysis; D007782:Lactoglobulins / Q000032:analysis* / Q000276:immunology; D008892:Milk / Q000737:chemistry / Q000276:immunology; D016269:Milk Hypersensitivity / Q000175:diagnosis* / Q000276:immunology; D011485:Protein Binding; D011817:Rabbits
chemical_list: D000485:Allergens; D000906:Antibodies; D000939:Epitopes; D007782:Lactoglobulins; D007073:Immunoglobulin E
keywords: Allergenic residues; Bovine β-lactoglobulin; ELISA; Milk allergens