重组戊型肝炎疫苗菌种库的建立及其稳定性  

Establishment and stability of seed bank of recombinant hepatitis E vaccine

作　　者：曹玉锋[1] 徐军[1] 时成波[1] 孟多佳[1] 迟春萍[1] 王玉梅[1] 贾媛[1] 张剑锋[1] 常东英[1] 王伟善[1] 

机构地区：[1]长春生物制品研究所有限责任公司生物技术室,长春130062

出　　处：《中国生物制品学杂志》2012年第11期1427-1430,共4页Chinese Journal of Biologicals

基　　金：吉林省科技发展项目(20080940);吉林省产业技术研究与开发项目(2009003)

摘　　要：目的建立重组戊型肝炎疫苗工程菌三级种子库,并分析其稳定性。方法将工程菌株HEV179/BL21(DE3)复苏活化,经全面检测合格后冻干保存即为原始种子库;原始种子库传代扩增,经全面检定合格后,冻干保存即为主种子库;主种子库经传代扩增和全面检定合格后,甘油冻存即为工作种子库。对三级种子库进行全面检定,并对工程菌进行冻干及甘油冻存稳定性、甘油冻存时间稳定性及质粒稳定性检测。结果建立的三级种子库经全面检定合格;菌种冻干稳定性、甘油冻存稳定性、甘油冻存时间稳定性及质粒稳定性指标均合格,菌种生长特性、遗传和表达稳定性均未发生变化。结论成功建立了HEV179/BL21(DE3)工程菌三级种子库,菌种稳定性良好,适用于重组戊型肝炎疫苗的工业化生产。Objective To establish three-tier seed bank of recombinant hepatitis E vaccine and study its stability. Methods Recombinant E. coli HEV179/BL21 （DE3） was resuscitated, activated and subjected to overall control tests, and the qualified ones were lyophilized as primary seed bank. The seeds from primary seed bank were subcultured, propagated and subjected to overall control tests, while the qualified ones were lyophilized as master seed bank. The seeds from master seed bank were subcuhured, propagated and subjected to overall tests, while the qualified ones were cryopreserved in glycerol and served as working seed bank. The three-tier seed bank was subjected to overall control tests, and the recombinant strain was evaluated for stabilities after lyophilization and cryopreservation in glycerol as well as the genetic stability. Results The established three-tier seed bank was qualified in overall control tests, of which the stabilities after lyophilization and cryopreservation in glycerol as well as the genetic stability met the relevant requirements. The growth characteristics as well as genetic and expression stabilities of recombinant strain showed no significant change. Conclusion Three-tier seed bank of HEV179/BL21 （DE3） was successfully established, and the seeds showed high stability and was suitable for industrial production of recombinant hepatitis E vaccine.

关 键 词：疫苗 肝炎病毒 戊型 种子库 稳定性 

分 类 号：R373.2[医药卫生—病原生物学] R392.33[医药卫生—基础医学]