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enGenes-X-press protein & DNA production platform
enGenes Biotech has developed X-press™ as an optimized solution for the cost-effective production of recombinant protein and plasmid DNA in microbial expression systems.Leveraged from enGenes’ world-class experience in cutting-edge genetic engineering, combined with a broad expertise in bioprocess engineering, the enGenes-X-press™ Escherichia coli (E.coli) proprietary technology forms a production platform that delivers substantially higher soluble product yields, easier downstream processing and up to 80% reduction in cost of goods sold (COGS).
The X-press technology platform allows recombinant product formation to be decoupled from cell growth, enabling bioprocessing with clear phase separation of biomass growth and product formation.
Using this procedure makes it possible to generate significantly higher specific and volumetric yields compared to the previous state-of-the-art (Escherichia coli BL21 (DE3).
Further, X-press developed bioprocesses are characterized by improved genetic stability, robustness and scalability.
The technology is based on a genetically modified host cell based on the Escherichia coli B-linage (BL21) that allows introduction of standard expression vectors (with T7 promoter) without cumbersome adaptations. The X-press fermentation process can be implemented with standard fed-batch fermentation equipment.
Reprogramming of the host cell is performed by co-expression of a bacteriophage-derived peptide that stops cell division and host mRNA production and at the same time modulates the host cell metabolism for improved soluble, high level protein production.
The technology also allows secretion of proteins targeted to the periplasmic space to the cell free supernatant, thereby allowing a cost-effective manufacturing option comparable to yeast-based expression systems.
The technology is fully scalable from lab to industrial scale (developed process running at >10 m³ scale).
enGenes-X-press example yields
The process advantages of enGenes X-press are borne out by sample protein and other yields:
- Green fluorescent protein (GFP) = 20 g/L
- Soluble enzyme (hydrolase) = 13 g/L (cytoplasm)
- Glycosyltransferase = 1 g/L (cytoplasm)
- Fab antibody fragment = 250 mg/L (cell supernatant)
- VHH (variable camelid heavy chain-only) single domain antibody = 1 g/L (cell supernatant)
- Affinity ligand = 8-10 g/L (cell supernatant)
The enGenes X-press protein production platform is thus made distinctive by a series of unique characteristics:
- E. coli based, growth decoupled production system
- Allowing decoupling of protein/DNA production from cell growth
- Host cell growth halted at defined cell concentration
- Protein of interest production triggered
- Host cell resources used exclusively for product of interest
- Proprietary IP (PCT/EP2016/059597)
X-press economic benefits
The economic benefits of the enGenes X-press platform extend into production and process efficiencies. They include:
- Higher product yields
- Significant yield improvement on previous E. coli BL21 (DE3) ‘gold standard’
- Enabling secretion of protein to cell supernatant
- Production of growth-interfering or toxic proteins/plasmids
- Improved genetic stability due to lack of cell division
- Continuous manufacturing possible
- Allowing improved Space/Time/Yield
enGenes-X-press forms a next-generation E. coli expression platform that allows for scalable production of difficult-to-express or even growth-interfering “toxic” recombinant proteins and plasmid DNA at a fraction of previous cost. Patents are pending on X-press technology as a growth-decoupled microbial protein production system.
Click on enGenes-X-press technology platform for more information.
Schematic of enGene-X-press process