重组乙型肝炎疫苗(酿酒酵母)中宿主细胞蛋白残留量检测方法的验证  

Validation of a detection method for residual host cell protein in recombinant hepatitis B vaccine(Saccharomyces cerevisiae)

作　　者：刘玉美 齐亭娟 张晓蕙 丁玲 陈若兰 栗妍 陈国明 柳易程 黄祎 Liu Yumei;Qi Tingjuan;Zhang Xiaohui;Ding Ling;Chen Ruolan;Li Yan;Chen Guoming;Liu Yicheng;Huang Yi(Quality Control Department,Beijing Institute of Biological Products Co.,Ltd.,Beijing 100176,China)

机构地区：[1]北京生物制品研究所有限责任公司质量检定室,北京100176

出　　处：《国际生物制品学杂志》2025年第1期46-49,共4页International Journal of Biologicals

摘　　要：目的验证重组乙型肝炎(乙肝)疫苗(酿酒酵母)中宿主细胞蛋白残留量ELISA定量检测方法。方法使用S.cerevisiae HCP ELISA Kit检测试剂盒,对重组乙肝疫苗(酿酒酵母)中的宿主细胞蛋白残留量进行检测,并验证该方法的线性、准确度、重复性、定量限、专属性及中间精密度。结果ELISA检测标准品的曲线线性良好(决定系数≥0.97),最低点吸光度(A_(405/492))值<0.300,最高点A_(405/492)值>1.000;同一检测人员对低、中、高3个浓度的加标样品进行3次重复检测,检测回收率在80%~120%;对中浓度样品重复检测6次,样品回收率在80%~120%,相对标准偏差<10%,表明方法具有良好的准确度和重复性。样品背景溶液和样品稀释液的A_(405/492)值均小于曲线最低点,表明该方法的专属性强;定量限为2 ng/mL;不同实验组在不同时间对中浓度样品重复检测6次,相对标准偏差<10%,中间精密度良好。结论ELISA检测重组乙肝疫苗(酿酒酵母)宿主细胞蛋白残留量具有良好的线性、准确度、重复性、定量限、专属性及中间精密度,可用于乙肝疫苗生产的过程控制。Objective To validate a quantitative ELISA method for the detection of residual host cell protein(HCP)in recombinant hepatitis B vaccine(Saccharomyces cerevisiae).Methods The S.cerevisiae HCP ELISA Kit was used to detect HCP residues in recombinant hepatitis B vaccine(Saccharomyces cerevisiae),and the linearity,accuracy,repeatability,limit of quantification,specificity and intermediate precision of the method were verified.Results The ELISA standard curve had good linearity(coefficient of determination≥0.97),with the lowest absorbance(A_(405/492))value<0.300,and the highest A_(405/492)value>1.000.With same experimenter,samples added with low,medium and high concentration standards were detected 3 times,and the recovery rates were 80%-120%.Medium concentration standard-added sample was detected 6 times,and the relative standard deviation(RSD)was<10%,indicating good accuracy and repeatability of the method.The A_(405/492)values of the sample background solution and the sample diluent were both smaller than the lowest point of the curve,indicating that the method was highly specific.The medium concentration standard-added sample was detected by different experimenter groups at different time for 6 times,and the RSD was<10%,indicating good intermediate precision of the method.Conclusion The ELISA method for the detection of HCP residues in recombinant hepatitis B vaccine(Saccharomyces cerevisiae)has good linearity,accuracy,repeatability,limit of quantification,specificity and intermediate precision,thus can be used for the process control of hepatitis B vaccine production.

关 键 词：酿酒酵母 蛋白残留量 检测 酶联免疫吸附测定 

分 类 号：R446.6[医药卫生—诊断学]