title: Development of an enzyme-linked immunosorbent assay system for detecting β'-component (Onk k 5), a major IgE-binding protein in salmon roe.
abstract: 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 0.78 μg/g and 2.60 μg/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.
authors: Faculty of Fisheries Sciences, Hokkaido University, Minato 3, Hakodate, Hokkaido 041-8611, Japan.
authors: Yutaka
authors: Y
authors: Shimizu
authors: Central Research Institute, Maruha Nichiro Corporation, Wadai 16, Tsukuba, Ibaraki 300-4295, Japan.
authors: Hiroshi
authors: H
authors: Oda
authors: Central Research Institute, Maruha Nichiro Corporation, Wadai 16, Tsukuba, Ibaraki 300-4295, Japan.
authors: Kohsuke
authors: K
authors: Seiki
authors: Faculty of Fisheries Sciences, Hokkaido University, Minato 3, Hakodate, Hokkaido 041-8611, Japan. Electronic address: saeki@fish.hokudai.ac.jp.
authors: Hiroki
authors: H
authors: Saeki
mesh_terms: D000485:Allergens / Q000032:analysis*; D000818:Animals; D004531:Eggs / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D029941:Fish Proteins / Q000032:analysis*; D005506:Food Contamination / Q000032:analysis; D005511:Food Handling; D037502:Galectin 3 / Q000032:analysis*; D008297:Male; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D012474:Salmon; D012680:Sensitivity and Specificity
chemical_list: D000485:Allergens; D029941:Fish Proteins; D037502:Galectin 3; C000593244:Onc k 5 allergen, salmon
keywords: ELISA; Enzyme immunoassay; Food allergy; Salmon roe; Yolk protein; β′-Component