利用牛奶过敏者血清检验牛奶中过敏原酶解程度的研究

Study on Determining the Degree of Enzymatic Hydrolysis of Allergen in Milk with Blood Serum of Allergic People to Milk

利用对牛奶过敏者和对牛奶不过敏者的血清作为“一抗”,采用酶联免疫吸附实验(ELISA)的方法,检测牛奶水解后对牛奶过敏者血清的结合能力,从而确定牛奶中过敏原蛋白的最佳水解消除条件,降低牛奶及奶制品对牛奶过敏者的致敏性,达到牛奶过敏人群可以饮用的标准。结果表明:胃蛋白酶、胰蛋白酶和木瓜蛋白酶共同水解牛奶的最佳条件是酶与底物质量比W(E/S)=4%,pH1.5(胃蛋白酶)和pH8.0(胰蛋白酶和木瓜蛋白酶),温度40℃(胃蛋白酶和胰蛋白酶)和45℃(木瓜蛋白酶),水解时间3h,用酶标仪在492nm测得其水解后奶样与过敏者血清结合的OD值已降到了牛奶不过敏者血清与未水解牛奶结合的OD值水平,达到了消除过敏原的效果。

The research used human blood serum allergic to milk and non-allergic to milk and applied ELISA method to determine the binding ability between human serum and milk hydrolyzed by enzyme. The optimum conditions of hydrolyzing milk and decreasing the danger of people＇s allergy and meeting the drinkable standards for human allergic to milk and dairy products were studied. The results indicated the optimum hydrolysis conditions were W（E/S）=4%, pH 1.5（pepsin）and pH8.0（trypsin and papain）, T=40℃（pepsin and trypsin） and 45℃（papain）, t=3h. The OD value at 492nm of the binding between the hydrolyzed milk and the blood serum of allergic people to milk decreased to the OD value of the binding between the non-hydrolyzed milk and the blood serum of non-allergic people to milk. On these conditions the allergen was eliminated.