Development of an enzyme-linked immunosorbent assay system for detecting β′-component (Onk k 5), a major IgE-binding protein in salmon roe

• Published in: Food chemistry Vol. 181; pp. 310 - 317
• Main Authors: Shimizu, Yutaka, Oda, Hiroshi, Seiki, Kohsuke, Saeki, Hiroki
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 15.08.2015
• Subjects: Allergens - analysis; Animals; Eggs - analysis; ELISA; Enzyme immunoassay; Enzyme-Linked Immunosorbent Assay - methods; Fish Proteins - analysis; Food allergy; Food Contamination - analysis; Food Handling; Galectin 3 - analysis; Male; Rabbits; Rats; Rats, Wistar; Salmon; Salmon roe; Sensitivity and Specificity; Yolk protein; β′-Component; Food allergy; β′-Component; Salmon roe; Yolk protein; Enzyme immunoassay; ELISA
• ISSN: 0308-8146; 1873-7072
• DOI: 10.1016/j.foodchem.2015.02.071

•We developed ELISA system for detecting β′-component (Onk k 5), a major IgE-binding protein in salmon roe.•Rabbit and rat polyclonal IgGs to β′-component were used for the development.•Protein extraction using SDS and 2-mercaptoethanol ensured specificity of ELISA.•The detection sensitivity of the developed ELISA system reached a practical level. A novel enzyme-linked immunosorbent assay (ELISA) system has been established for selective detection of chum salmon (Oncorhynchus keta) yolk protein (SYP). Rabbit and rat polyclonal Immunoglobulin G antibodies to β′-component (the major allergic protein in fish roe; anti-β) were applied for designing the ELISA system. The sandwich ELISA using rabbit anti-β for the capture antibody and horseradish peroxidase-labeled F(ab′)2 fragment of rat anti-β for the detection antibody obtained high sensitivity and narrow specificity for SYP. Protein extraction using sodium dodecyl sulfate and 2-mercaptoethanol ensured strict specificity of the ELISA, and components of three popular processed foods had no effect on the ELISA response. The limits of determination and quantification of SYP were estimated to be 0.78μg/g and 2.60μg/g of food sample, respectively. In conclusion, the developed ELISA system has a probability to be applied for the detection of contaminated chum salmon roe in processed food.