本文選題：須癬毛癬菌 + 細(xì)胞壁蛋白��； 參考：《中國免疫學(xué)雜志》2017年09期

【摘要】：目的:提取須癬毛癬菌的細(xì)胞壁蛋白作為免疫原制備特異性卵黃抗體,并鑒定其生物活性,為卵黃抗體在預(yù)防與治療皮膚癬疾病的應(yīng)用奠定基礎(chǔ)。方法:本研究采用冷堿抽提的方法提取須癬毛癬菌的細(xì)胞壁蛋白,并用其免疫健康的產(chǎn)蛋母雞。采用聚乙二醇兩步沉淀法及飽和硫酸銨鹽析提純卵黃抗體;用Bradford法檢測卵黃抗體中蛋白含量;SDS-PAGE凝膠電泳分析測定卵黃抗體的純度以及相對分子質(zhì)量;ELISA檢測純化的卵黃抗體的效價;Western blot分析特異性卵黃抗體的免疫反應(yīng)性。結(jié)果:提取的卵黃抗體中蛋白純度達(dá)到87.27%。由細(xì)胞壁蛋白制備的特異性卵黃抗體在初免20 d后效價開始升高,到45天達(dá)到最高值(1∶32 000)。Western blot結(jié)果顯示由細(xì)胞壁蛋白制備的卵黃抗體能與其良好的特異性結(jié)合,具有較好的免疫反應(yīng)性。結(jié)論:本實驗提取的須癬毛癬菌細(xì)胞壁蛋白作為免疫原可以制備出特異性較強(qiáng)的卵黃抗體,為須癬毛癬菌感染的皮膚疾病提供了治療新思路。
[Abstract]:Objective: to prepare the specific yolk antibody by extracting the cell wall protein of Trichophyton tubuloides as immunogen, and to identify its biological activity, so as to lay a foundation for the application of yolk antibody in the prevention and treatment of dermatophytosis. Methods: the cell wall protein of Trichophyton tinea was extracted by cold alkali extraction, and the healthy laying hen was immunized with it. The yolk antibody was purified by means of polyethylene glycol two-step precipitation and saturated ammonium sulfate salting-out. The protein content in yolk antibody was detected by Bradford method. The purity of yolk antibody was determined by SDS-PAGE gel electrophoresis. The titer of purified egg yolk antibody was detected by relative molecular weight Elisa. Western blot was used to analyze the immunoreactivity of specific yolk antibody. Results: the purity of yolk antibody was 87.27%. The titer of the specific yolk antibody prepared from the cell wall protein began to increase 20 days after the first immunization, and reached its highest value at 45 days (1:32 000). Western blot results showed that the yolk antibody prepared from the cell wall protein could bind to the yolk antibody with good specificity. It has good immunoreactivity. Conclusion: the cell wall protein extracted in this experiment can be used as immunogen to prepare specific egg yolk antibody, which provides a new idea for the treatment of dermatophytes infected by Trichophyton tubuloides.
【作者單位】： 廣東工業(yè)大學(xué)輕工化工學(xué)院;
【分類號】：R756

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