食品中のタンパク質性アレルゲンの迅速同定法

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The determination of specific serum IgE has been evaluated for the diagnosis of allergy. Although it is possible to detect the proteins capable of binding the serum IgE in vitro by RAST, Western blot and ELISA, the determined protein is not always shown to be a genuine allergen of the allergy in progress. Therefore, it is necessary for the patients to get correct information of allergens. In the present study, we established a method to identify the true allergen within various proteins contained in foods. The concept of this method is based on the principle that an anaphylactic reaction could be induced with any of the proteins in the sensitizing food separated in SDS-PAGE (sodium dodecylsulfate-polyacrylamide gel electrophoresis), if the mouse was sensitized with a certain food. Thus, this method indicative of anaphylactic reaction makes it possible to identify the allergen just cross-linking the IgEs on mast cells in vivo. In this paper, we used hen egg-white lysozyme (HEL) as a model antigen to confirm this method. Normal male ddY mice of 5 weeks of age were sensitized intraperitoneally with HEL emulsified in Freund's incomplete adjuvant (FIA). Nine to 14 days after the sensitization, antigen challenge in abdominal wall was performed in two different forms. After SDS-PAGE, the one was an injection of homogenized segments of acrylamide gel containing HEL, and the other was an injection of HEL eluted from gel segments by electrophoresis or buffer. It was possible to induce the HEL specific anaphylactic reaction by both challenges. These results demonstrated that a small amount of ingredient separated by SDS-PAGE, even in the presence of the gel, could be applied to AW method. Hence, by the present study, the advantage of SDS-PAGE, in addition to the specification of proteins binding to IgE (Western blot), is expanded to the confirmation whether the protein is true allergen of anaphylactic reaction. The extensive application of this method to identify the true allergen in foods that contains varieties of protein is expected.
日本食品化学学会の論文
2001-08-31

食品中のタンパク質性アレルゲンの迅速同定法

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