牛奶主要致敏原组分的分离纯化和分析

发布时间：2018-04-28 23:39

本文选题：食品过敏原 + 牛奶　；参考：《暨南大学》2008年硕士论文

【摘要】： [目的]牛奶过敏症的诊断治疗以及低/无过敏原配方奶研制的关键是牛奶过敏原组分的分离、纯化及标准化。本研究分离纯化出牛奶中引起过敏反应的致敏原组分,通过分析与组胺释放的关系研究其免疫活性进而鉴定出牛奶主要过敏原组分,为研制适用于中国人的过敏原标准化疫苗提供依据。 [方法]采用等电点沉淀、(NH_4)_2SO_4盐析法、Sephadex G-50凝胶层析和DEAE-Sepharose离子交换层析等方法分离、纯化牛奶过敏原组分。分别以脱脂乳、乳清、酪蛋白和β-Lg免疫C57/BL6小鼠。免疫印迹分析牛奶致敏原组分。间接ELISA检测免疫小鼠血清中sIgE和sIgG;同时改良荧光法检测牛奶不同致敏原组分体外诱发96孔板内不同免疫原腹腔致敏C57/BL6小鼠MC组胺释放率。 [结果]分离、纯化出酪蛋白、β-Lg、α-La等牛奶过敏原组分,纯度＞90%。免疫印迹表明酪蛋白是牛奶主要致敏原组分。间接ELISA检测免疫C57/BL6小鼠血清中sIgE与sIgG;结果表明抗血清sIgG水平显著升高,sIgE水平低;组胺释放率与sIgG水平是一致的,而与sIgE水平没有相关性。脱脂乳中最主要的过敏原组分为酪蛋白,乳清中最主要的过敏原组分为β-Lg。组胺释放实验结果表明酪蛋白和β-Lg之间有重要的相似表位。 [结论]组胺荧光检测法和sIgG水平可用于牛奶过敏原组分的鉴定和评价,为食物过敏原标准化提供研究方法。
[Abstract]:[objective] the key to the diagnosis and treatment of milk hypersensitivity and the development of low / no allergen formula is the separation, purification and standardization of milk allergen. In this study, the allergen components of milk were isolated and purified, and the main allergen components of milk were identified by analyzing the relationship between the hypersensitivity and histamine release. To provide the basis for the development of Chinese allergen standardized vaccine. [methods] Sephadex G-50 gel chromatography and DEAE-Sepharose ion exchange chromatography were used to purify the allergen from milk. C57/BL6 mice were immunized with skim milk, whey, casein and 尾-LG respectively. The allergen components of milk were analyzed by Western blot. Indirect ELISA was used to detect sIgE and Siga in serum of immunized mice, and modified fluorescence method was used to detect the release rate of MC histamine in 96-well intraperitoneal sensitized C57/BL6 mice induced by different allergen components of milk in vitro. [results] casein, 尾 -Lg, 伪 -La and other milk allergens were isolated and purified. Western blot showed that casein was the main allergen of milk. Indirect ELISA was used to detect the levels of sIgE and Siga in serum of C57/BL6 mice. The results showed that the level of sIgG in antiserum was significantly higher than that of sIgE, and the histamine release rate was consistent with the level of sIgG, but had no correlation with the level of sIgE. Casein is the main allergen in skim milk and 尾 -Lg is the main allergen in whey. Histamine release assay showed that casein and 尾-LG had important similar epitopes. [conclusion] histamine fluorescence assay and sIgG level can be used to identify and evaluate the components of milk allergen and provide a method for the standardization of food allergens.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

【参考文献】