首批Vero细胞蛋白质标准品的研制

Preparation of the first batch of Vero cell protein standard

目的制备首批Vero细胞蛋白质国家标准品,用于人用狂犬病疫苗Vero细胞宿主残留量检测。方法采用Vero细胞分泌蛋白质为原料,加入蔗糖冻干制成Vero细胞蛋白质国家标准品。本标准品的赋值由两步完成:第一步采用Lowry法对Vero细胞蛋白质标准品原液进行蛋白含量测定;第二步采用酶联免疫法以Vero细胞蛋白质标准品原液绘制标准曲线对标准品进行蛋白含量赋值。随机选取16支蛋白质标准品,测定pH值,采用方差分析统计检验其均匀性;随机选取标准品,分别在不同温度(4、25、37℃)下放置不同时间(1、2、3和4周),各时间点取3支,酶联免疫法检测抗原含量,采用方差分析统计检验其稳定性。结果经协作标定,Vero细胞蛋白质标准品的标示量为2.6μg/mL,标准偏差为0.4,95%可信区间为2.51~2.78μg/mL。该批标准品的均匀性及稳定性良好。结论目前该Vero细胞蛋白质标准品已获批使用,批号为250022-201901,该标准品的建立,对提高人用狂犬病疫苗的质量控制具有重要意义。

Objective To prepare the first batch of Vero cell protein standard for detection of residual Vero cell host pro-tein in rabies vaccine for human use.Methods Vero cell protein standard was prepared by using Vero cell secretory protein as the raw material and sucrose as the protective agent for lyophilization,of which the assignment was completed by two steps.The first step was the determination of protein content in bulk of Vero cell protein standard by Lowry method,while the second step was the assignment of protein content of Vero cell protein standard by ELISA method using the standard curve plotted with the bulk of the standard.Sixteen containers of Vero cell protein standard were randomly selected and determined for pH value to test the uniformity by analysis of variance.The standard was randomly selected and stored at various temperatures(4,25 and 37℃)for various times durations(1,2,3 and 4 weeks),from which three containers were taken at each time point and determined for antigen content by ELISA to test the stability by analysis of variance.Results Collaborative calibration showed that the stated amount of Vero cell protein standard was 2.6μg/mL,with a standard deviation of 0.4μg/mL and a 95%confidence interval of 2.51~2.78μg/mL.This batch of standard showed good uniformity and stability.Conclusion At present,Vero cell protein standard has been approved for use,of which batch number is 250022-201901.The establishment of this standard is of great significance in improving the quality control of rabies vaccine for human use.