Geisse Sabine, Fux Cornelia
Novartis Institutes for BioMedical Research, Department of NBC/PPA, Basel, Switzerland.
Methods Enzymol. 2009;463:223-38. doi: 10.1016/S0076-6879(09)63015-9.
The timely availability of recombinant proteins in sufficient quantity and of validated quality is of utmost importance in driving drug discovery and the development of low molecular weight compounds, as well as for biotherapeutics. Transient gene expression (TGE) in mammalian cells has emerged as a promising technology for protein generation over the past decade as TGE meets all the prerequisites with respect to quantity and quality of the product as well as cost-effectiveness and speed of the process. Optimized protocols have been developed for both HEK293 and CHO cell lines which allow protein production at any desired scale up to >100 l and in milligram to gram quantities. Along with an overview on current scientific and technological knowledge, detailed protocols for expression of recombinant proteins on small, medium, and large scale are discussed in the following chapter.
在推动药物发现、低分子量化合物开发以及生物治疗方面，及时获得足量且经过验证质量的重组蛋白至关重要。在过去十年中，哺乳动物细胞中的瞬时基因表达（TGE）已成为一种有前景的蛋白质生产技术，因为TGE在产品数量和质量、成本效益以及生产过程速度方面均满足所有先决条件。已针对HEK293和CHO细胞系开发了优化方案，可实现高达>100升的任何所需规模的蛋白质生产，产量可达毫克至克级。在下一章中，除了对当前科学技术知识进行概述外，还将讨论重组蛋白在小、中、大规模表达的详细方案。
