摘要
目的研制用于人用狂犬病疫苗(地鼠肾细胞)中地鼠肾细胞蛋白质残留量检测(ELISA法)的国家标准品。方法分两步进行协作标定和量值溯源,第一步采用Lowry法对原代地鼠肾细胞蛋白质标准品原液进行赋值,第二步采用ELISA法对原代地鼠肾细胞蛋白质标准品进行赋值。随机抽取16支标准品,检测pH值,采用方差分析统计检验均匀性;将标准品分别在4、25和37℃放置不同时间(1、2、3和4周),不同时间点取3支,于-20℃保存,待所有样本放置结束后统一用ELISA法检测蛋白含量,方差分析统计检验稳定性。结果协作标定的数据经统计学分析,原代地鼠肾细胞蛋白质标准品赋值为4.0μg/mL,95%可信限为(4.0±0.5)μg/mL,均匀性及稳定性良好。结论建立了国家原代地鼠肾细胞蛋白质标准品(批号:250024-201901),赋值准确可靠、科学严谨,对人用狂犬病疫苗(地鼠肾细胞)的质量控制具有重要意义。
Objective To develop the national standard for determination of residual hamster kidney cells in rabies vaccine(hamster kidney cells)for human use by ELISA method.Methods Cooperative calibration and trace of measures were performed by two steps.In the first step,Lowry method was used to assign the bulk of primary hamster kidney cell protein standard.However,in the second step,ELISA method was used to assign the primary hamster kidney cell protein standard.Sixteen containers of standard were randomly selected and determined for pH value,of which the uniformity was verified by analysis of variance.The standard was stored at 4,25 and 37℃for various time durations(1,2,3 and4 weeks),from which three containers were taken at each time points and determined for protein content by ELISA,and the stability was verified by analysis of variance.Results According to the results of statistical processing of data in cooperative calibration,the content of standard for primary hamster kidney cell protein was assigned as 4.0μg/mL,of which the 95%confidence limit was(4.0±0.5)μg/mL.The standard also showed good uniformity and stability.Conclusion National standard for primary hamster kidney cell protein was established(Lot:250024-201901),of which the assigned value was accurate,reliable,scientific and rigorous.It is of an important significance in quality control of rabies vaccine(hamster kidney cells).
作者
石磊泰
李加
张月兰
王辉
雷继军
郭中平
李玉华
SHI Lei-tai;LI Jia;ZHANG Yue-lan;WANG Hui;LEI Ji-jun;GUO Zhong-ping;LI Yu-hua(National Institutes for Food and Drug Control,Beijing 102629,China)
出处
《中国生物制品学杂志》
CAS
CSCD
2020年第10期1186-1191,共6页
Chinese Journal of Biologicals
基金
国家药典委员会药品标准提高课题“疫苗细胞基质残余蛋白测定方法的建立”资助。
