细胞工厂制备F基因型腮腺炎减毒活疫苗工艺初步研究  

Preliminary study on the production process for F-genotype attenuated mumps vaccine in cell factory

作　　者：杨振清 邱恒 王微[1] 郑俊杰 赵卫中[1] 何云龙 李琦涵[1] 王晶晶[1] Yang Zhenqing;Qiu Heng;Wang Wei;Zheng Junjie;Zhao Weizhong;He Yunlong;Li Qihan;Wang Jingjing(Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Diseases,Institute of Medical Biology,Chinese Academy of Medicine Science&Peking Union Medical College,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：《国际生物制品学杂志》2020年第6期282-286,共5页International Journal of Biologicals

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

摘　　要：目的:初步建立以人二倍体细胞KMB 17为培养基质的F基因型腮腺炎减毒活疫苗细胞工厂工艺。方法:分别使用细胞工厂和细胞培养瓶,以含体积分数5%和10%牛血清的培养基将KMB 17细胞培养至单层后计数,并按不同感染复数(multiplicity of infection,MOI)接种F基因型腮腺炎病毒,观察细胞病变情况,初步确定细胞工厂工艺最适MOI和病毒收获时间。在无血清培养基条件下,对两种生产工艺制备的腮腺炎减毒活疫苗半成品和成品进行病毒滴度检测、热稳定性试验及其他相关质量指标的检测。结果:在细胞工厂中获得了生长状态良好的KMB 17细胞,经洗涤后,在MOI为0.020时,培养7~9 d后可获得滴度较高的病毒收获液,经过滤、冻干,成品病毒滴度较稳定,其他质量指标经检测均符合中国药典2015年版三部的要求。结论:初步建立了细胞工厂制备F基因型腮腺炎减毒活疫苗的生产工艺。Objective To preliminarily establish the cell factory production process of F-genotype attenuated mumps vaccine in human diploid cell KMB17.Methods Cell factory and cell culture bottle were used to culture KMB17 cells to monolayer with 5%and 10%bovine serum,respectively.Cells were counted,inoculated with F-genotype mumps virus at different multiplicities of infection(MOIs)in serum-free culture medium.The cytopathic effect was observed after virus inoculation to determine the most suitable MOI and harvest time.The virus titer,thermal stability and other related quality items of bulk and final products were tested for 2 different production processes in serum-free culture medium.Results KMB17 cells obtained from cell factory were in good growth condition.After cell wash,with MOI of 0.020,virus harvests with high virus titer were obtained after 7-9 d of culture.After filtration and freeze-drying,virus titer of the final product was stable,and other quality items were in line with the requirements of Chinese pharmacopoeia 2015(VolumeⅢ).Conclusion The production process of F-genotype attenuated mumps vaccine in human diploid cell KMB17 is preliminarily established.

关 键 词：腮腺炎疫苗 细胞工厂 KMB 17细胞 

分 类 号：R392-33[医药卫生—免疫学]