本文選題：食品過敏原 + 牛奶��； 參考：《暨南大學(xué)》2008年碩士論文

【摘要】： [目的]牛奶過敏癥的診斷治療以及低/無(wú)過敏原配方奶研制的關(guān)鍵是牛奶過敏原組分的分離、純化及標(biāo)準(zhǔn)化。本研究分離純化出牛奶中引起過敏反應(yīng)的致敏原組分,通過分析與組胺釋放的關(guān)系研究其免疫活性進(jìn)而鑒定出牛奶主要過敏原組分,為研制適用于中國(guó)人的過敏原標(biāo)準(zhǔn)化疫苗提供依據(jù)。 [方法]采用等電點(diǎn)沉淀、(NH_4)_2SO_4鹽析法、Sephadex G-50凝膠層析和DEAE-Sepharose離子交換層析等方法分離、純化牛奶過敏原組分。分別以脫脂乳、乳清、酪蛋白和β-Lg免疫C57/BL6小鼠。免疫印跡分析牛奶致敏原組分。間接ELISA檢測(cè)免疫小鼠血清中sIgE和sIgG;同時(shí)改良熒光法檢測(cè)牛奶不同致敏原組分體外誘發(fā)96孔板內(nèi)不同免疫原腹腔致敏C57/BL6小鼠MC組胺釋放率。 [結(jié)果]分離、純化出酪蛋白、β-Lg、α-La等牛奶過敏原組分,純度＞90%。免疫印跡表明酪蛋白是牛奶主要致敏原組分。間接ELISA檢測(cè)免疫C57/BL6小鼠血清中sIgE與sIgG;結(jié)果表明抗血清sIgG水平顯著升高,sIgE水平低;組胺釋放率與sIgG水平是一致的,而與sIgE水平?jīng)]有相關(guān)性。脫脂乳中最主要的過敏原組分為酪蛋白,乳清中最主要的過敏原組分為β-Lg。組胺釋放實(shí)驗(yàn)結(jié)果表明酪蛋白和β-Lg之間有重要的相似表位。 [結(jié)論]組胺熒光檢測(cè)法和sIgG水平可用于牛奶過敏原組分的鑒定和評(píng)價(jià),為食物過敏原標(biāo)準(zhǔn)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, 偽 -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.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392

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