ArticlePloS one2013
Multi-host expression system for recombinant production of challenging proteins.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Response Surface Methodology to Optimize the Expression Efficiency of Recombinant Reteplase.Iranian journal of biotechnology · 2023Article
- Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment.Viruses · 2022Article
- In vivo enzymatic digestion of HRV 3C protease cleavage sites-containing proteins produced in a silkworm-baculovirus expression system.Bioscience reports · 2022Article
- Phage Display-Derived Compounds Displace hACE2 from Its Complex with SARS-CoV-2 Spike Protein.Biomedicines · 2022Article
- The short isoform of the host antiviral protein ZAP acts as an inhibitor of SARS-CoV-2 programmed ribosomal frameshifting.Nature communications · 2021Article
- Live-Cell Cardiac-Specific High-Throughput Screening Platform for Drug-Like Molecules that Enhance CaCells · 2020Article
- A drug-inducible sex-separation technique for insects.Nature communications · 2020Article
- ER intrabody-mediated inhibition of interferon α secretion by mouse macrophages and dendritic cells.PloS one · 2019Article
- Identifying parameters to improve the reproducibility of transient gene expression in High Five cells.PloS one · 2019Article
- Fast plasmid based protein expression analysis in insect cells using an automated SplitGFP screen.Biotechnology and bioengineering · 2016Article
- Algal autolysate medium to label proteins for NMR in mammalian cells.Journal of biomolecular NMR · 2016Article
- Identification of Essential Genetic Baculoviral Elements for Recombinant Protein Expression by Transactivation in Sf21 Insect Cells.PloS one · 2016Article
- Efficient Mammalian Cell Expression and Single-step Purification of Extracellular Glycoproteins for Crystallization.Journal of visualized experiments : JoVE · 2015Article
- Cellular disulfide bond formation in bioactive peptides and proteins.International journal of molecular sciences · 2015Review
- Genomic Analysis and Isolation of RNA Polymerase II Dependent Promoters from Spodoptera frugiperda.PloS one · 2015Article
- Polyethyleneimine-based transient gene expression processes for suspension-adapted HEK-293E and CHO-DG44 cells.Protein expression and purification · 2013Article
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6 authors.
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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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