摘要
探究细胞培养生产流感病毒过程中,高细胞密度引起低单位细胞病毒产率(Svy)的原因及找到解决方法。通过增加微载体浓度以及换液操作获得较高的细胞密度;考察了感染时细胞密度(CCI)和病毒感染用维持培养基对病毒产量和单位细胞病毒产率的影响。在12.6 g/L微载体培养过程中,通过换液操作,可获得高细胞密度,达到1.47×107 cells/m L。在CCI为1×107cells/m L条件时,选择合适的维持培养基,其Svy最高达到5.14×103 virions/cell,比同等条件下用DMEM维持培养基的Svy值提高了近一倍。高密度培养MDCK细胞生产流感病毒时,充分考虑不同阶段的培养条件和营养需求,可以提高细胞密度和单位细胞病毒产率,进而提高流感病毒的产量。该研究结果为工业化生产流感病毒疫苗提供了基础数据。
It was to investigate the phenomenon that high cell density with low cell-specific virus yields and find out the solution. Methods：High cell density was achieved by increasing the microcarrier concentration and enhancing nutrition supply, then the effect of cell concentration at infection（CCI）and virus maintenance medium（VMM）on virus titer and cell-specific virus yield（Svy）was studied. Results：The maximum cell density was up to 1.47＆#215;107cells/mL in 12.6 g/L microcarrier culture using medium exchange strategy. Under high CCI（1＆#215;107cells/mL）condition, the Svy was up to 5.14＆#215;103 virions/cell by choosing appropriate virus maintenance medium, which is nearly twice of Svy in DMEM. Conclusion：In order to achieve higher influenza virus titer, culture conditions and nutrition demands at different stages of influenza production should be taken into consideration. The results in this work provide guidance for further development of industrial-scale vaccine production processes.
出处
《生物技术通报》
CAS
CSCD
北大核心
2015年第1期203-208,共6页
Biotechnology Bulletin
基金
国家自然科学基金项目(21106045
21206040)
国家"863"计划(2012AA02A303)
国家重大专项(2013ZX10004003-003-003)
中央高校基本科研业务费(WF1214035)
关键词
高密度培养
MDCK
流感
病毒
high-density culture
MDCK
influenza production
virions
