卵黄抗体三种提取方法的比较研究

Comparison of Extracting Egg Yolk Antibody by Three Different Methods

关键词:  卵黄抗体  提取  硫酸铵法  聚乙二醇法  冷乙醇法

Key words:  egg yolk antibody  extraction  ammonium sulfatemethod  PEG method  cold ethanol method

摘要:

运用硫酸铵法、聚乙二醇（PEG）法、冷乙醇法对卵黄抗体进行提取，并使用间接竞争酶联免疫吸附试验（ELISA）方法测定所得卵黄抗体的特异性，用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）测其纯度，用Nanodrop2000测其蛋白浓度并计算提取量，比较三种方法所得抗体的特异性、纯度和提取量。结果显示，三种提取方法所得卵黄抗体特异性高低依次为硫酸铵法、冷乙醇法、PEG法；纯度高低依次为硫酸铵法、冷乙醇法、PEG法；三种方法的提取量高低依次为PEG法、硫酸铵法、冷乙醇法。经综合分析与评价，硫酸铵法的抗体特异性较好、纯度较高、提取量也较高，且其操作方法相对简便，是比较具有优势的提取方法。

Abstract:

In our research, egg yolk antibody was extracted by ammonium sulfate method, polyethylene glycol method (PEG) method and cold ethanol method, and its specificity, purity, and the extract quantity were determined by indirect competitive enzyme-linked immunosorbent assay(ELISA) method, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method and Nanodrop2000, respectively. The result showed that the specificity and purity of egg yolk extracted by ammonium sulfatemethod was better than that of cold ethanol method which seemed better than PEG method; however, the extracting ratio of egg yolk antibody extracted by three methods from high to low was that PEG method, ammonium sulfate method and cold ethanolmethod. In summary, antibody extracted by the ammonium sulfate method presented good specificity, high purity and ideal extracting ratio with the relatively simple operation, so ithad more advantages than other two methods.