无血清细胞培养在发热伴血小板减少综合征布尼亚病毒生长中的应用  

Application of serum free cell culture in production of severe fever with thrombocytopenia syndrome bunyavirus

作　　者：许定花 王建梅 周鹏 杨小青 冼嘉仪 XU Ding-hua;WANG Jian-mei;ZHOU Peng;YANG Xiao-qing;XIAN Jia-yi(Research and Development Department, Wuxi Sinosbio Biomedical Technologies, Wuxi 214000, Jiangsu Province, China)

机构地区：[1]无锡鑫连鑫生物医药科技有限公司研发部,江苏无锡214000

出　　处：《微生物学免疫学进展》2018年第2期5-8,共4页Progress In Microbiology and Immunology

基　　金：863计划(2014AA021003)

摘　　要：目的建立无血清培养基培养Vero细胞制备发热伴血小板减少综合征布尼亚病毒(severe fever with thrombocytopenia syndrome bunyavirus,SFTSV)的工艺。方法分别采用含10%牛血清的MEM(10%MEM培养基)和无血清M2培养基(SF-M2培养基)在方瓶中培养Vero细胞制备SFTSV,比较无血清与含血清培养基培养Vero细胞制备SFTSV在病毒滴度及病毒繁殖曲线之间的差异。在生物反应器里用无血清培养的方式进行工艺放大,收获病毒原液并进行检定。结果无血清培养的Vero细胞能够满足SFTSV培养需求,与含血清细胞培养相比,单位细胞病毒产量没有降低,达到30~60个活病毒/细胞。可以实现在生物反应器的工艺放大,病毒高峰时病毒滴度均在7.0lg PFU/m L以上。结论无血清细胞培养可以应用于SFTSV的培养,有利于降低疫苗生产过程中的纯化难度,提高疫苗安全性。Objective To develop a procedure in preparation of severe fever with thrombocytopenia syndrome bunyavirus （SFTSV） by culture of Vero cells in serum-free medium. Methods SFTSV was inoculated into Vero cells cultured in MEM containing 10% bovine serum （10% MEM culture） and M2 （SF-M2 culture） medium without serum, respectively. Virus titer and proliferation curve of SFTSV were detected and compared in both conditions. Amplification was carried out in ser- um-free cultivation of Vero cell in a bioreactor, the virus was harvested and detected. Results Compared with serum-con- taining medium, Vero cells cultured by serum-free medium could meet the requirements of SFTSV production without cell reduction, and reached to a yield about 30-60 live viruses/cell. The scale-up process cultured by serum-free medium could be achieved in the bioreactor with the virus titers 37.0 IgPFU/mL on the peak of virus production. Conclusion Serum- free medium could be applied in cultivation of SFTSV, which is of benefit to reduce the difficulty in virus purification and to improve the safety of the vaccine.

关 键 词：无血清细胞培养 发热伴血小板减少综合征布尼亚病毒 生物反应器 安全性 

分 类 号：R392-33[医药卫生—免疫学]