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昆虫细胞在14升气升式生物反应器的无血清培养基中生产重组β-半乳糖苷酶。

Recombinant beta-galactosidase production in serum-free medium by insect cells in a 14-L airlift bioreactor.

作者信息

King G A, Daugulis A J, Faulkner P, Goosen M F

机构信息

Department of Chemical Engineering, Queen's University, Kingston, Ontario, Canada.

出版信息

Biotechnol Prog. 1992 Nov-Dec;8(6):567-71. doi: 10.1021/bp00018a015.

Abstract

Spodoptera frugiperda (Sf9) insect cells were successfully cultured in serum-free medium in a 14-L airlift bioreactor. Cell densities as high as 1 x 10(7) cells/mL were achieved with specific growth rates of approximately 0.0286 h-1 (doubling time of 24 h). This system was also used to demonstrate the expression of a reported gene, beta-galactosidase (beta-gal), when cells were infected with a recombinant baculovirus. Approximately 0.33 mg of beta-gal/mL (i.e., 104,000 units/mL) of medium were obtained at the 14-L scale, while about 0.95 mg of beta-gal/mL (i.e., 285,000 units/mL) of medium were obtained in small-scale shaker flasks. The difference was attributed to a suboptimal infection in the large scale. Specific oxygen consumption rates decreased from 5.58 x 10(-17) mol O2/cell.s in early exponential growth to 3.13 x 10(-17) mol O2/cell.s at 3 days post-infection.

摘要

草地贪夜蛾(Sf9)昆虫细胞在14升气升式生物反应器的无血清培养基中成功培养。细胞密度高达1×10⁷个细胞/毫升,比生长速率约为0.0286 h⁻¹(倍增时间为24小时)。当细胞被重组杆状病毒感染时,该系统还用于证明报告基因β-半乳糖苷酶(β-gal)的表达。在14升规模下,每毫升培养基获得约0.33毫克β-gal(即104,000单位/毫升),而在小规模摇瓶中每毫升培养基获得约0.95毫克β-gal(即285,000单位/毫升)。差异归因于大规模培养中感染不理想。比耗氧率从指数生长早期的5.58×10⁻¹⁷摩尔O₂/细胞·秒降至感染后3天的3.13×10⁻¹⁷摩尔O₂/细胞·秒。

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