Determination of Crustacean Allergen in Food Products by Sandwich ELISA

DOI Web Site 12 Citations 10 References
  • Shibahara Yusuke
    Research Division, Nissui Pharmaceutical Co., Ltd.
  • Oka Michihiro
    Research Division, Nissui Pharmaceutical Co., Ltd.
  • Tominaga Kei
    Research Division, Nissui Pharmaceutical Co., Ltd.
  • Ii Toshihiro
    Research Division, Nissui Pharmaceutical Co., Ltd.
  • Umeda Mamoru
    Research Division, Nissui Pharmaceutical Co., Ltd.
  • Uneo Noriko
    Food Safety Research Center, Nippon Suisan Kaisha, Ltd.
  • Abe Akihisa
    Food Safety Research Center, Nippon Suisan Kaisha, Ltd.
  • Ohashi Eiji
    Food Safety Research Center, Nippon Suisan Kaisha, Ltd.
  • Ushio Hideki
    Faculty of Marine Science, Tokyo University of Marine Science and Technology
  • Shiomi Kazuo
    Faculty of Marine Science, Tokyo University of Marine Science and Technology

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Other Title
  • サンドイッチＥＬＩＳＡ法による食品中の甲殻類アレルゲンの検出
  • サンドイッチ ELISAホウ ニ ヨル ショクヒンチュウ ノ コウカクルイ アレルゲン ノ ケンシュツ

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Crustaceans are one of the most significant sources of food allergies, and tropomyosin is their major allergen. To detect crustacean allergens in food products, we have developed a sandwich ELISA using a crustacean tropomyosin-specific monoclonal antibody using black tiger prawn tropomyosin as an immunogen. Our ELISA showed 82-102% reactivity (based on reactivity to black tiger prawn tropomyosin) to tropomyosins from crustaceans including prawns, lobster, crab, hermit crab, and krill, while no substantial reactivity (<0.1%) to those from mollusks, including squid, octopus, and shellfish, was observed. Furthermore, no positive ELISA results were obtained with any crustacean-free foods. Repeatability coefficients of variation (CVs) were <10%, and the detection limit based on a standard crustacean protein was 0.16ppm, which qualified for the sensitivity requirements of the current food labeling system. The ELISA could detect crustacean tropomyosin, even heat-denatured tropomyosin, without being affected by food matrices. These results indicated that this ELISA is highly specific to crustacean tropomyosin, and will be useful as a method for detecting crustacean allergens in processed food products.

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