Multi-host expression system for recombinant production of challenging proteins.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
MetadataShow full item record
AbstractRecombinant 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.
CitationMulti-host expression system for recombinant production of challenging proteins. 2013, 8 (7):e68674 PLoS ONE
AffiliationDepartment of Molecular Structural Biology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
The following license files are associated with this item:
- High-level recombinant protein production in CHO cells using an adenoviral vector and the cumate gene-switch.
- Authors: Gaillet B, Gilbert R, Amziani R, Guilbault C, Gadoury C, Caron AW, Mullick A, Garnier A, Massie B
- Issue date: 2007 Jan-Feb
- A comparative study of different vector designs for the mammalian expression of recombinant IgG antibodies.
- Authors: Li J, Menzel C, Meier D, Zhang C, Dübel S, Jostock T
- Issue date: 2007 Jan 10
- High-level recombinant protein production in CHO cells using lentiviral vectors and the cumate gene-switch.
- Authors: Gaillet B, Gilbert R, Broussau S, Pilotte A, Malenfant F, Mullick A, Garnier A, Massie B
- Issue date: 2010 Jun 1
- Auditioning of CHO host cell lines using the artificial chromosome expression (ACE) technology.
- Authors: Kennard ML, Goosney DL, Monteith D, Roe S, Fischer D, Mott J
- Issue date: 2009 Oct 15
- Enhancement of recombinant human EPO production and glycosylation in serum-free suspension culture of CHO cells through expression and supplementation of 30Kc19.
- Authors: Park JH, Wang Z, Jeong HJ, Park HH, Kim BG, Tan WS, Choi SS, Park TH
- Issue date: 2012 Nov