鸡胚细胞疫苗制品宿主细胞蛋白残留量检测方法的建立

Development of an ELISA kit for determination of residual chick embryo cell proteins in vaccines

目的建立鸡胚细胞疫苗制品中宿主细胞蛋白残留量的检测方法。方法制备鸡胚细胞蛋白和抗鸡胚细胞蛋白抗体，摸索ELISA双抗体夹心法的试验条件，建立酶联免疫检测方法，并进行各项指标的验证。结果纯化的鸡胚细胞蛋白抗体纯度达到90％以上，抗体效价可达1：128。蛋白质印迹法分析显示，纯化抗体与鸡胚细胞蛋白呈现特异性结合。该方法的线性范围为12．5—200．0ng／ml，相关系数≥0．9885；对已知浓度样品的回收率为92％～110％，变异系数为3．0％-9．2％。结论建立的ELISA双抗体夹心法对于检测鸡胚细胞疫苗制品中残留的蛋白含量具有良好的线性、专属性、精密性及准确性。

Objective To develop an ELISA kit for determination of residual chick embryo cell proteins in vaccines. Methods Chick embryo cell proteins and their corresponding antibodies were prepared and used for development of an ELISA kit. A standard curve was plotted with determination results of the prepared reference and various indexes of the developed kit were tested and verified. Results The purity and titer of the antibodies purified from immunized rabbits reached. 〉 90 % and 1： 128, respectively. Western blot showed specific binding of the purified antibodies with most of the proteins. The chick embryo cell protein reference at concentrations of 12.5-200.0 ng/ml showed a good linear relationship （r≥0.9885）. The recovery rates of known concentration samples were 92%-110% and coefficients of variation were 3. 0%-9. 2%. Conclusions The developed ELISA kit for residual chick embryo cell proteins in vaccines shows good linear relationship, specificity, precision and accuracy.