Fertig G, Rahn H P, Angermann A, Klöppinger M, Miltenburger H G
Institute of Zoology, Technical University of Darmstadt, Germany.
Cytotechnology. 1993;11(1):67-75. doi: 10.1007/BF00749059.
We have investigated large scale production processes (up to 2 liters) of recombinant proteins using the baculovirus expression system in order to optimize the product yields. Experiments using cell lines of Spodoptera frugiperda (Sf9) and Mamestra brassicae (IZD-Mb0503) were performed to show the different production capacities of the cell lines. The influence of the infection at different cell densities is described. Beyond that, TC100-, IPL41- and serum-free IPL41-medium were compared to demonstrate their different capabilities of supporting cell growth and protein expression. Additionally, the inhibitory effect of FCS on the protease activity of kallikrein, which is produced in its zymogenic form, is discussed Improved production parameters are described, which enabled us to produce up to 8000 units of activated pro-kallikrein within 14 days using perfusion cultivation.
我们研究了使用杆状病毒表达系统大规模生产重组蛋白(高达2升)的过程,以优化产品产量。使用草地贪夜蛾(Sf9)和甘蓝夜蛾(IZD-Mb0503)的细胞系进行实验,以展示细胞系的不同生产能力。描述了不同细胞密度下感染的影响。除此之外,还比较了TC100培养基、IPL41培养基和无血清IPL41培养基,以证明它们支持细胞生长和蛋白质表达的不同能力。此外,还讨论了胎牛血清(FCS)对以酶原形式产生的激肽释放酶蛋白酶活性的抑制作用。描述了改进的生产参数,这些参数使我们能够在14天内使用灌注培养法生产多达8000单位的活化前激肽释放酶。