免疫斑点印迹法检测水解乳蛋白婴幼儿配方粉免疫反应  

Detection of the Immunoreaction of Hydrolyzed Milk Protein in Infant Formula Using Dot-immunobinding Blot Assess

作　　者：孙丽娟[1] 韩诗雯 曾冰蕙 段素芳 屠振华 李放 车会莲[1] Sun Lijuan;Han Shiwen;Zeng Binghui;Duan Sufang;Tu Zhenhua;Li Fang;Che Huilian(College of Food Science and Nutritional Engineering,China Agriculture University,Beijing 100083;Inner Mongolia Yili Industrial Group Co.,Ltd.,Inner Mongolia Dairy Technology Research Institute Co.,Ltd.,Hohhot 010110;China National Research Institute of Food&Fermentation Industries Co.,Ltd.,Beijing 100015)

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083 [2]内蒙古伊利实业集团股份有限公司内蒙古乳业技术研究院有限责任公司,呼和浩特010110 [3]中国食品发酵工业研究院有限公司,北京100015

出　　处：《中国食品学报》2024年第7期342-350,共9页Journal of Chinese Institute Of Food Science and Technology

基　　金：中国食品科学技术学会食品科技基金-伊利健康科学基金项目(2022-Y02);国家自然科学基金面上项目(31972185)。

摘　　要：目的:建立一种快速且精准的方法,以确定水解乳蛋白婴幼儿配方粉的免疫反应性。方法:首先采用直接免疫斑点印迹法,利用牛奶过敏患儿血清初步评估待测样品的免疫反应性;然后通过间接竞争免疫斑点印迹法以市售普通牛奶粉为阳性对照,评估水解乳蛋白婴幼儿配方粉与血清免疫球蛋白E(IgE)的结合能力;最后采用间接和间接竞争酶联免疫检测法,以佐证免疫斑点印迹结果。结果:部分水解乳蛋白婴配粉仍可以结合IgE以诱发过敏反应,而深度水解乳蛋白婴配粉不具有免疫反应性。免疫斑点印迹和酶联免疫检测评价免疫反应性结果显著且一致。结论:免疫斑点印迹技术操作性强,周期短,成本低,结果可靠,为利用较难获取的婴幼儿过敏患儿血清初步评估乳蛋白产品致敏性提供了新思路。Objective:Cow's milk allergy(CMA)is the most common type of food allergy in infants and young children.Infants with milk allergy need to use low-allergenic hydrolyzed cow milk protein formula for the prevention and management.Although protein allergen levels can be reduced through enzymatic hydrolysis,depending on the type of enzymes used and the various filtering methods,residual antigenic epitopes can still cause food allergies.Therefore,the purpose of this work was to develop a quick and accurate method to assess the immunoreactivity of hydrolyzed cow's milk protein infant formula powder.Methods:Firstly,a direct dot-immunobinding blot assay was used to gauge the samples'immunoreactivity with serum samples from allergic children.Subsequently,an indirect competitive dot-immunobinding blot assay was utilized,using commercially available unhydrolyzed whole protein milk powder as a positive control and other milk product samples as competitive antigens,to evaluate the binding capacity of hydrolyzed milk protein infant powder to serum immunoglobulin E(IgE).Finally,direct,and indirect competitive enzyme-linked immunosorbent assay(ELISA)were also used to validate and confirm the results.Results:The study's findings demonstrated that partially hydrolyzed formula can still bind IgE to induce allergic reactions,while extensively hydrolyzed formula is not immunoreactive.This study provides a new approach for the preliminarily evaluation of the allergenicity of milk protein products using limited serum samples from infants and young children with allergies.

关 键 词：牛奶过敏 免疫斑点印迹 酶联免疫吸附 间接和间接竞争 过敏血清 

分 类 号：TS252.51[轻工技术与工程—农产品加工及贮藏工程]