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Multi-host expression system for recombinant production of challenging proteins.
Meyer, Steffen Lorenz, Carmen Baser, Bahar Wördehoff, Mona Jäger, Volker van den Heuvel, Joop
Meyer, Steffen
Lorenz, Carmen
Baser, Bahar
Wördehoff, Mona
Jäger, Volker
van den Heuvel, Joop
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Abstract
Recombinant production of complex eukaryotic proteins for structural analyses typically requires a profound screening process to identify suitable constructs for the expression of ample amounts of properly folded protein. Furthermore, the evaluation of an optimal expression host has a major impact on protein yield and quality as well as on actual cost of the production process. Here we present a novel fast expression system for multiple hosts based on a single donor vector termed pFlp-Bac-to-Mam. The range of applications of pFlp-Bac-to-Mam comprises highly efficient transient transfection of HEK293-6E in serum-free suspension culture and subsequent large-scale production of challenging proteins expressing in mg per Liter level using either the baculoviral expression vector system or stable CHO production cell lines generated by Flp-mediated cassette exchange. The success of the multi-host expression vector to identify the optimal expression strategy for efficient production of high quality protein is demonstrated in a comparative expression study of three model proteins representing different protein classes: intracellular expression using a fluorescent protein, secretion of a single-chain-Fv-hIgG1Fc fusion construct and production of a large amount of highly homogeneous protein sample of the extracellular domain of a Toll-like receptor. The evaluation of the production efficiency shows that the pFlp-Bac-to-Mam system allows a fast and individual optimization of the expression strategy for each protein class.
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Multi-host expression system for recombinant production of challenging proteins. 2013, 8 (7):e68674 PLoS ONE
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en
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1932-6203
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