抗须癣毛癣菌细胞壁蛋白的卵黄抗体制备和鉴定

发布时间：2018-06-29 00:39

本文选题：须癣毛癣菌 + 细胞壁蛋白　；参考：《中国免疫学杂志》2017年09期

【摘要】：目的:提取须癣毛癣菌的细胞壁蛋白作为免疫原制备特异性卵黄抗体,并鉴定其生物活性,为卵黄抗体在预防与治疗皮肤癣疾病的应用奠定基础。方法:本研究采用冷碱抽提的方法提取须癣毛癣菌的细胞壁蛋白,并用其免疫健康的产蛋母鸡。采用聚乙二醇两步沉淀法及饱和硫酸铵盐析提纯卵黄抗体;用Bradford法检测卵黄抗体中蛋白含量;SDS-PAGE凝胶电泳分析测定卵黄抗体的纯度以及相对分子质量;ELISA检测纯化的卵黄抗体的效价;Western blot分析特异性卵黄抗体的免疫反应性。结果:提取的卵黄抗体中蛋白纯度达到87.27%。由细胞壁蛋白制备的特异性卵黄抗体在初免20 d后效价开始升高,到45天达到最高值(1∶32 000)。Western blot结果显示由细胞壁蛋白制备的卵黄抗体能与其良好的特异性结合,具有较好的免疫反应性。结论:本实验提取的须癣毛癣菌细胞壁蛋白作为免疫原可以制备出特异性较强的卵黄抗体,为须癣毛癣菌感染的皮肤疾病提供了治疗新思路。
[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.
【作者单位】：广东工业大学轻工化工学院;
【分类号】：R756

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