重组杆状病毒感染悬浮昆虫细胞感染策略优化及扩大试验  

Optimization of infection strategy for recombinant baculovirus infection of suspended insect cells and its scale-up test

作　　者：张佑红[1] 靖志强[1] 马静[1] 徐鹏[1] 陈龙[1] 

机构地区：[1]武汉工程大学化工与制药学院绿色化工过程省部共建教育部重点实验室,湖北武汉430073

出　　处：《化工学报》2010年第4期942-948,共7页CIESC Journal

基　　金：国家自然科学基金项目(20876120);武汉市科技攻关项目(20052002044)~~

摘　　要：昆虫细胞-杆状病毒表达系统已经广泛应用于生物杀虫剂及重组蛋白的大规模生产。在昆虫细胞-杆状病毒表达系统中,感染策略中的感染复数(MOI)、感染时间(TOI)、细胞初始浓度(ICD)的相互关系还不是很清楚,而且相关报道也很少。本研究在摇瓶中利用正交实验探讨了MOI、TOI及ICD的相互作用关系以及对包涵体病毒(OV)和非包涵体病毒(BV)产量的影响。结果表明,感染复数对OV和BV产量有显著的影响;最佳感染条件为MOI为0.1,TOI为细胞处于对数生长期初期,ICD为2×105cells.ml-1;最大BV病毒滴度为2.0×107TCID50.ml-1。将在摇瓶中获得的最佳条件在气升式反应器进行扩大试验,获得最大病毒滴度为2.13×107TCID50.ml-1。Insect cell-baculovirus expression system has been widely used in insect pesticide and recombinant protein production on a large scale.The interplay among the multiplicity of infection（MOI）,time of infection（TOI）and initial cell density（ICD）in insect cell-recombinant expression systems is not clearly understood and the literature related to the investigation of the quantitative relationship of MOI,TOI and ICD is very scarce.In the present study,an orthogonal design was used to select the optimum conditions,in which the effects of MOI,ICD and TOI on the budded virus（BV）titer and occluded virus（OV）number were evaluated in the HzAM1 insect cell-recombinant baculovirus system at a low MOI in shake-flasks.The results indicated that among MOI,TOI and ICD,MOI had a significant influence on the amounts of BVs and OVs produced.The optimal conditions for the production of both BVs and OVs in the HzAM1 insect cell-recombinant baculovirus system were：MOI of 0.1,initial cell density of 2×105 cells·ml-1 and infection at the early exponential phase.A maximum BV concentration of 2.0×107 TCID50·ml-1,was obtained.Furthermore,the optimal condition obtained was applied to the scale-up test in the airlift reactor,and a maximum BV concentration of 2.13×107 TCID50·ml-1 was obtained,which was bigger than those in the shaked flask.

关 键 词：昆虫杀虫剂 杆状病毒 昆虫细胞 感染策略 

分 类 号：Q819[生物学—生物工程]