牛奶过敏原制备方法的研究和免疫活性鉴别

Investigation on the preparation of milk allergens and the identification of immunoreactivity

目的探索牛奶主要过敏原的制备工艺。方法采用等电点沉淀、凝胶层析和分子筛等技术纯化牛奶中主要过敏原组分;采用SDS-PAGE鉴定蛋白纯度,采用双抗原夹心-ELISA鉴定免疫活性。结果成功获得牛奶中的四种主要过敏原组分:酪蛋白、β乳球蛋白、α乳白蛋白和分子量较高的P1组分,并经过免疫实验证实这四种组分均能与过敏血清产生反应,而其中β乳球蛋白和P1组分反应性较高。结论本研究探索出一种简单、实用的牛奶主要过敏原制备工艺,并证实牛奶中的β乳球蛋白和高分子量蛋白质为主要过敏原。

Objective To investigate the method of preparing main components of milk allergens. Methods Isoelectric precipitation, gel chromatography, molecular sieve and other possible methods were used to purify the main allergens in milk. SDS-PAGE was used to identify the purity of protein and double antigen sandwich-EL1SA was established to confirm the immunoreactivity of milk allergen. Results Four components of milk allergen, including caseins, 13-1actoglobulin, alactoalbumin and several macromolecule proteins （ P1 ） , were separated and purified by the methods established in this study, and all these components can react with positive serums, whereas f3-1actoglobulin and P1 components showed higher immunoreactivity. Conclusion A simple and practical method for preparing milk allergens was found in this study; J3- laetoglobulin and PI components were considered to be the main allergen components in milk.