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。
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 cold ethanol extracted by specificity and purity of egg yolk extracted by ammonium sulfatemethod was better than that of method which seemed better three methods from high than PEG method; however, the extracting ratio of egg yolk antibody 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.
出处
《中国兽药杂志》
北大核心
2015年第8期51-54,共4页
Chinese Journal of Veterinary Drug
关键词
卵黄抗体
提取
硫酸铵法
聚乙二醇法
冷乙醇法
egg yolk antibody
extraction
ammonium sulfatemethod
PEG method
cold ethanol method
