A time- and cost-efficient system for high-level protein production in mammalian cells
Acta Crystallographica Section D
Volume 62, Issue 10, pages 1243–1250, October 2006
How to Cite
Aricescu, A. R., Lu, W. and Jones, E. Y. (2006), A time- and cost-efficient system for high-level protein production in mammalian cells. Acta Crystallographica Section D, 62: 1243–1250. doi: 10.1107/S0907444906029799
- eukaryotic expression;
- mammalian cells;
- protein production.
Most proteins for structural biology studies are produced by high-level expression in Escherichia coli. However, prokaryotic based expression systems fail to generate correctly folded functional forms of many proteins and hence a variety of eukaryotic based expression systems have been developed. Of these, yeast and baculovirus-infected insect cells currently represent the expression systems of choice for structural biologists. Here, protocols for a simple, fast and affordable method for transient protein expression in mammalian cells are reported. The results demonstrate that it combines several features necessary for the production of suitable samples for structural biology, in particular protein crystallography, namely high protein yield, straightforward purification, selenomethionine incorporation and control of N-linked glycosylation. The system is suitable for use in conventional laboratories or can be implemented in a medium- or high-throughput pipeline.