title: A sensitive enzyme-linked immunosorbent assay for the determination of fish protein in processed foods.
abstract: Fish is one of the most common causes of food allergy and its major allergen is parvalbumin, a 12 kDa muscular protein. In this study, a sandwich enzyme-linked immunosorbent assay (ELISA) for the determination of fish protein in processed foods was developed using a polyclonal antibody raised against Pacific mackerel parvalbumin. The developed sandwich ELISA showed 22.6-99.0% reactivity (based on the reactivity to Pacific mackerel parvalbumin) to parvalbumins from various species of fish. The limits of detection and quantitation were estimated to be 0.23 and 0.70 μg protein per g of food, respectively. When the sandwich ELISA was subjected to inter-laboratory validation, spiked fish protein was recovered from five model processed foods in the range of 69.4-84.8% and the repeatability and reproducibility relative standard deviations were satisfactorily low (≤ 10.5%). Thus, the sandwich ELISA was judged to be a useful tool to determine fish protein in processed foods.
authors.affiliation: Nissui Pharmaceutical Co. Ltd., Research Division, 1075-2 Hokunanmoro, Yuki-shi, Ibaraki 307 0036, Japan. y-shibahara@yki.nissui-pharm.jp
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Shibahara
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Uesaka
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: Shoichi
authors.initials: S
authors.lastname: Yamada
authors.forename: Kazuo
authors.initials: K
authors.lastname: Shiomi
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D057140:Fast Foods / Q000032:analysis*; D029941:Fish Proteins / Q000032:analysis*; D005399:Fishes; D012680:Sensitivity and Specificity
chemical_list: D029941:Fish Proteins