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昆虫细胞(Bm5)悬浮培养中重组蛋白生产的最佳感染条件。

Optimum infection conditions for recombinant protein production in insect cell (Bm5) suspension culture.

作者信息

Zhang J, Kalogerakis N, Behie L A, Iatrou K

机构信息

Faculty of Engineering, University of Calgary, Alberta, Canada.

出版信息

Biotechnol Prog. 1994 Nov-Dec;10(6):636-43. doi: 10.1021/bp00030a008.

Abstract

The baculovirus/insect cell expression system is an efficient and practical method for the production of many active therapeutic proteins on a large scale. The advantages of suspension cultures have been demonstrated with the study of a baculovirus/insect cell (BmNPV/Bm5) expression system for the production of recombinant chloramphenicol acetyltransferase (CAT), a model heterologous protein. Key infection parameters such as infection time and multiplicity of infection were examined systematically for the maximization of protein production. Furthermore, emphasis was placed on the development of possible medium replenishment strategies, which were necessary to achieve higher volumetric protein production from the infection of high-density cell cultures without sacrificing specific protein productivity. The highest protein production was achieved with the infection of suspended cells in the mid to late exponential growth phase.

摘要

杆状病毒/昆虫细胞表达系统是一种高效且实用的大规模生产多种活性治疗性蛋白质的方法。通过研究杆状病毒/昆虫细胞(BmNPV/Bm5)表达系统生产重组氯霉素乙酰转移酶(CAT,一种典型的异源蛋白),已证明了悬浮培养的优势。系统地研究了关键感染参数,如感染时间和感染复数,以实现蛋白质产量最大化。此外,重点在于开发可能的培养基补充策略,这对于在不牺牲特定蛋白质生产率的情况下,通过高密度细胞培养感染实现更高的体积蛋白质产量是必要的。在指数生长中期至后期感染悬浮细胞时,实现了最高的蛋白质产量。

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