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建立甲型肝炎灭活疫苗制品中Vero细胞残留蛋白含量测定方法。制备Vero细胞蛋白抗原,免疫家兔制备抗血清,提取纯化抗血清IgG并以酶标记,建立酶联免疫吸附法检测系统。用Vero细胞蛋白抗原参考品绘制定量标准曲线,并对各项指标进行验证。结果显示兔抗IgG效价为1:32,通过Western blot表明兔抗IgG与Vero细胞蛋白抗原能特异性结合;通过方阵试验确定抗体包被浓度为1:400,酶标抗体工作浓度为1:200～1:400,抗原参考品检测范围为25～800ng/ml,线性关系良好,r=0.995,该试剂特异性、精密度及准确度良好,稳定性试验证明在4℃可保存半年。建立了酶联免疫吸附法,可用于甲型肝炎灭活疫苗中Vero细胞残留蛋白含量的检测。
To establish a method for determination of Vero cell residual protein content in hepatitis A inactivated vaccine preparations. Vero cell protein antigens were prepared, rabbits were immunized to prepare antiserum, purified anti-serum IgG and labeled with enzyme to establish enzyme-linked immunosorbent assay system. Quantitative standard curve was drawn with Vero cell protein antigen reference, and each index was verified. The results showed that the rabbit anti-IgG titer was 1:32, Western blot showed that rabbit anti-IgG and Vero cell protein antigen specific binding; by matrix test to determine the antibody coating concentration of 1: 400, ELISA antibody working concentration of 1 : 200 ~ 1: 400. The detection range of antigen reference was 25 ~ 800ng / ml, the linearity was good, r = 0.995. The specificity, precision and accuracy of this reagent were good. The stability test proved that it could be stored for half a year at 4 ℃. The enzyme-linked immunosorbent assay (ELISA) was established to detect the residual protein content of Vero cells in hepatitis A inactivated vaccine.