Backliwal Gaurav, Hildinger Markus, Chenuet Sebastien, Dejesus Maria, Wurm Florian M
Ecole Polytechnique Fédérale de Lausanne, Laboratory of Cellular Biotechnology, Institute of Bioengineering, Faculty of Life Sciences, Lausanne, Switzerland.
N Biotechnol. 2008 Oct-Dec;25(2-3):162-6. doi: 10.1016/j.nbt.2008.08.007. Epub 2008 Aug 27.
Recombinant proteins are of great commercial and scientific interest. However, most current production methods using mammalian cells involve the time- and labor-intensive step of creating stable cell lines. Although production methods based on transient gene expression could offer a significant improvement, transient transfection is currently still limited by low titers and low specific productivity compared to stable cell lines. To overcome these bottlenecks, we have explored the use of various growth factors to enhance specific productivity and titers in the context of transient gene expression. For that purpose, several growth factors were cloned and screened for their effect on transient gene expression in HEK293E and CHO-DG44 cells. In particular, acidic fibroblast growth factor (aFGF) was able to increase specific productivity by 60% and recombinant protein titers by 80% in HEK293E cells, while FGF9 increased titers by 250% in CHO-DG44 cells.
重组蛋白具有重大的商业和科学价值。然而，目前大多数使用哺乳动物细胞的生产方法都涉及创建稳定细胞系这一耗时且费力的步骤。尽管基于瞬时基因表达的生产方法可能会带来显著改进，但与稳定细胞系相比，瞬时转染目前仍受限于低滴度和低比生产率。为了克服这些瓶颈，我们探索了在瞬时基因表达的背景下使用各种生长因子来提高比生产率和滴度。为此，克隆了几种生长因子，并筛选它们对HEK293E和CHO - DG44细胞中瞬时基因表达的影响。特别是，酸性成纤维细胞生长因子（aFGF）能够使HEK293E细胞的比生产率提高60%，重组蛋白滴度提高80%，而FGF9能使CHO - DG44细胞的滴度提高250%。
