重组乙型肝炎疫苗(CHO细胞)冻干参考品的研制

Preparation of freeze-dried recombinant hepatitis B vaccine(CHO cells) reference

目的制备重组乙型肝炎疫苗(CHO细胞)冻干参考品,用于重组乙型肝炎疫苗(CHO细胞)的效力评价。方法选取检定合格的重组乙型肝炎疫苗(CHO细胞)原液,经协作标定表面抗原蛋白含量后,加入氢氧化铝佐剂和冻干保护剂,冷冻干燥制备重组乙型肝炎疫苗(CHO细胞)冻干参考品,并按《中国药典》三部(2010版)要求进行各项检定。经10次独立试验测定冻干参考品小鼠效力,采用Reed-Münch计算ED50值;将冻干参考品置4℃(8周)、37℃(4和8周)后分别检测疫苗效力,分析其稳定性,依据Q10法进行效期推测;并对2个生产企业10批疫苗进行效力测定,分析其适用性。结果制备的冻干参考品各项指标均符合规定,小鼠ED50均值为0.183μg,CV为50.5%;该CHO细胞效力冻干参考品蛋白含量定为20μg/ml,规格为10μg/0.5 ml;冻干保护剂、冻干工艺对乙型肝炎疫苗(CHO细胞)的效力影响较小,冻干参考品在37℃放置不同时间,效力变化不明显,表明冻干参考品稳定性较好,有效期约为7年;10批疫苗中,不合格率为10%,表明该参考品可用于乙型肝炎疫苗(CHO细胞)效力质控。结论制备的冻干参考品可作为重组乙型肝炎疫苗(CHO细胞)效力检定的质控参考品。

Objective To prepare freeze-dried recombinant hepatitis B vaccine（CHO cells） reference for evaluation of potency of recombinant hepatitis B vaccine（CHO cells）.Methods Bulk of qualified recombinant hepatitis B vaccine（CHO cells） was selected as the raw material of reference.After collaborative calibration of HBsAg content,the bulk was added with aluminium adjuvant and preservative,then lyophilized to prepared freeze-dried recombinant hepatitis B vaccine reference on which overall control tests were performed according to the requirements in Chinese Pharmacopoeia（Volume Ⅲ,2010 edition）.The potency of the freeze-dried reference was determined by ten independent tests in mice,and the ED50 value was calculated by Reed-Münch method.The freeze-dried reference was stored at 4 ℃ for 8 weeks and 37 ℃ for 4 and 8 weeks,and determined for potency,based on which the stability was analyzed,and the validity period was predicted by Q10 method.Ten batches of vaccine manufactured by two manufacturers were determined for potency and analyzed for suitability.Results All the quality indexes of prepared reference met the relevant requirements,of which the mean ED50 value to mice was 0.183 μg,with a CV value of 50.5%.The protein content of the reference was 20 μg / ml,while the specification was 10 μg / 0.5 ml.Neither preservative nor lyophilization procedure showed significant influence on the potency.After storage at 37 ℃ for various weeks,the potency of reference showed no significant change,indicating high stability.The validity period of the reference was about 7 years.Of the ten batches of vaccines,10% were unqualified,indicating that the reference was suitable for the potency control of hepatitis B vaccine（CHO cells）.Conclusion The prepared freeze-dried reference may be used for potency control of recombinant hepatitis B vaccine（CHO cells）.