CAJ | 학술논문

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2×10~5 cells·ml~(-1); 最大BV病毒滴度为2.0×10~7 TCID_(50)·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2.13×10~7 TCID_(50)·ml~(-1).
곤충세포-간상병독표체계통이경엄범응용우생물살충제급중조단백적대규모생산.재곤충세포-간상병독표체계통중,감염책략중적감염복수(MOI)、감염시간(TOI)、세포초시농도 (ICD)적상호관계환불시흔청초,이차상관보도야흔소.본연구재요병중이용정교실험탐토료MOI、TOI급ICD적상호작용관계이급대포함체병독(OV)화비포함체병독(BV)산량적영향.결과표명,감염복수대OV화BV산량유현저적영향; 최가감염조건위MOI위0.1,TOI위세포처우대수생장기초기,ICD위2×10~5 cells·ml~(-1); 최대BV병독적도위2.0×10~7 TCID_(50)·ml~(-1).장재요병중획득적최가조건재기승식반응기진행확대시험,획득최대병독적도위2.13×10~7 TCID_(50)·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 2×10~5 cells·ml~(-1) and infection at the early exponential phase.A maximum BV concentration of 2.0×10~7 TCID_(50)·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 2.13×10~7 TCID_(50)·ml~(-1) was obtained,which was bigger than those in the shaked flask.