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A time- and cost-efficient system for high-level protein production in mammalian cells

✍ Scribed by Aricescu, A. Radu ;Lu, Weixian ;Jones, E. Yvonne

Publisher
International Union of Crystallography
Year
2006
Tongue
English
Weight
517 KB
Volume
62
Category
Article
ISSN
0907-4449

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✦ Synopsis

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.

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