F基因型腮腺炎减毒活疫苗(人二倍体细胞)SP-A株工作种子批的建立与检定  

Establishment and quality control of working virus seeds of F-genotype attenuated mumps vaccine SP-A strain(human diploid cell)

作　　者：邱恒 田文志 王微[1] 郑俊杰 任芳芳[1] 段颖融 李琦涵[1] 王晶晶[1] QIU Heng;TIAN Wen-zhi;WANG Wei;ZHENG Jun-jie;REN Fang-fang;DUAN Ying-rong;LI Qi-han;WANG Jing-jing(Institute of Medical Biology,Chinese Academy of Medical Science&Peking Union Medical College,Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Diseases,Kunming 650118,China)

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所,云南省重大传染病疫苗研发重点实验室,昆明650118

出　　处：《中国新药杂志》2021年第15期1381-1385,共5页Chinese Journal of New Drugs

基　　金：中国医学科学院医学与健康科技创新工程资助项目(2016-12M-1-019);云南省科技人才和平台计划资助项目(2019HB073)。

摘　　要：目的:以人二倍体细胞KMB-17株为细胞培养基质建立F基因型腮腺炎减毒活疫苗SP-A株工作种子批。方法:在细胞工厂中以血清浓度为5%的MEM培养基培养KMB-17细胞,待细胞生长至单层后,分别按照0.020,0.050和0.100感染复数(multiplicity of infection,MOI)的比例接种病毒,逐日检测病毒滴度,绘制病毒增殖动力曲线以确定最适MOI和最佳收获时间。制备工作种子批,测定其病毒滴度及热稳定性,同时进行鉴别实验、无菌检查、分枝杆菌检查、支原体检查、病毒外源因子检查和小疏水蛋白(small hydrophobic protein,SH)核酸序列检查。利用该工作种子制备3批病毒收获液和疫苗原液并进行相关质量检定。结果:根据病毒增殖动力曲线,确定工作种子批的最适MOI为0.020,接种后d 7为病毒最佳收获时间,按此条件制备了工作种子批并完成检定,结果符合《中华人民共和国药典》2020年版三部腮腺炎减毒活疫苗毒种制造及检定质量要求。利用该工作种子批制备了3批病毒收获液和疫苗原液,经检定均符合《中华人民共和国药典》2020年版三部病毒收获液和原液的制造及检定规程要求。结论:建立F基因型腮腺炎减毒活疫苗SF-A株工作种子批,为后续F基因型腮腺炎减毒活疫苗的产业化提供了支持。Objective:To establish the working virus seeds of F-genotype attenuated mumps vaccine SP-A strain in human diploid cells KMB-17.Methods:In cell factory,KMB-17 cells were cultured in MEM medium with 5%newborn bovine serum.After the cells grew to monolayer,the virus was inoculated into 0.020,0.050 and 0.100 multiplicity of infection(MOI),respectively,in KMB-17 cells.The virus titer was detected everyday,and the optimal MOI and harvest time were determined from the dynamic proliferation curve of virus.The working virus seeds were prepared,the virus titer and thermal stability were determined.Identity,sterility,Mycobacterium Mycoplasma,Adventitious virus and small hydrophobic protein(SH)nucleic acid sequence test were carried out on the working virus seeds.Three batches of experimental vaccine virus harvest and bulk were prepared by using the working virus seeds,and relevant quality control was carried out.Results:According to the dynamic proliferation curve of virus,optimistic MOI was determined as 0.020,and the best harvest time was 7 days after inoculation.The working virus seeds were prepared according to the optimized procedure.The working virus seeds and three batches of experimental vaccine virus harvest and bulk met the requirements of"Chinese Pharmacopoeia"(VolumeⅢ,2020 edition).Conclusion:The results established procedure of working virus seeds of F-genotype attenuated mumps vaccine SP-A strain,and these data support the subsequent large-scaled preparation of F-genotype attenuated mumps vaccine.

关 键 词：腮腺炎疫苗 工作种子 生产工艺 检定 

分 类 号：R927[医药卫生—药学] R512.1