摘要
目的应用三种不同方法分离提取特异性抗人蔗糖酶卵黄抗体,比较总蛋白浓度、蛋白纯度及生物活性。方法以人蔗糖酶蛋白作为抗原免疫蛋鸡,分别采用硫酸钠水提取法、聚乙二醇法和冷乙醇法分离提取卵黄抗体,BCA法测定总蛋白质含量;聚丙烯酰胺凝胶电泳法检测抗体纯度;间接酶联免疫吸附测定法检测抗体效价。结果经典的硫酸钠水提取法相对其他两种方法提取的卵黄抗体在总蛋白含量和纯度都较高,杂带相应较少,聚乙二醇法提取的抗体效价较低,硫酸钠水提取法和冷乙醇法提取的抗体效价基本没有区别。结论三种提取方法中,硫酸钠水提取法操作简单、纯度较高、对蛋白影响较小,为后续有效分离提取卵黄抗体打下实验基础。
Objective Sucrase yolk antibodies acquired through three different separation and extraction methods, to compare total protein concentration, protein purity and biological activity. Methods Hens were injected by the purified sucrase. IgY was extracted by sodium sulfate water extraction methods, PEG6000 methods and EtOH. The protein content was detected by BCA kit. The protein purity was analyzed by SDS -PAGE ElectropHoresis, antibody titers was analyzed by indirect ELISA. Results That classic sodium sulfate water extraction method was better than the other two methods, with less Non - target bands compared to the other two methods. The antibodies extracted by cold EtOH method with low titers, titer of antibody extracted by sodium sulfate water extraction methods and PEG6000 methods without difference. Conclusions Sodium sulfate water extraction methods have the advantages including simple operation, high purity and less impact on protein among the three extracts methods, provide foundation for effective separation and extraction of yolk antibodies.
出处
《齐齐哈尔医学院学报》
2016年第19期2438-2441,共4页
Journal of Qiqihar Medical University
基金
贵州省科技厅基金项目(黔科合SY字(2012)3091)
遵义医学院青年科研启动基金资助(F-501)
关键词
蔗糖酶
卵黄抗体
分离提取
抗体纯度
生物活性
Sucrase
Egg yolk antibody
Separation and extraction
Antibody purity
Biological activity
