enGenes-e^Xchange™ technology for antibiotic resistance-free recombinant proteins and plasmid DNA

enGenes Biotech has developed -e^Xchange™ as an optimized solution for the cost-effective production of recombinant proteins and plasmid DNA that are free of antibiotic resistance genes.

It can be used in conjunction with other enGenes technology platforms, such as -e^Xpress™ and enGenes-e^Xtra™ for enhanced production of very high-quality plasmid DNA (pDNA) and recombinant proteins, allowing for faster-track production of enzymes and antigen development, for instance.

enGenes-e^Xpress™

Leveraged from enGenes’ world-class experience in cutting-edge genetic engineering, combined with a broad expertise in bioprocess engineering, the enGenes-e^Xpress™ Escherichia coli (E.coli) proprietary technology forms a production platform that delivers up to five times higher soluble product yields and easier downstream processing.

enGenes-eXpress™ is a patented next-generation, growth-decoupled Escherichia coli expression platform that enables scalable, cost-effective production of difficult-to-express or toxic recombinant proteins and plasmid DNA, reducing cost of goods sold (COGS) by up to 80%.

The technology is based on a genetically modified host cell variant of BL21 (DE3) that allows introduction of standard expression vectors (with T7 promoter) without cumbersome media adaptations. Using this procedure makes it possible to generate significantly higher specific and volumetric yields compared to the previous ‘gold standard’ E. coli BL21 (DE3).

enGenes-e^Xpress™ technology enables the growth-decoupled production of biomolecules using a modified form of the E. coli bioprocess. This is a versatile technology that can be used to produce various biomolecules, including recombinant proteins, plasmid DNA, RNA, and small molecules, but at higher productivity, with improved quality and reduced production costs. The T7 phage-derived inhibitor peptide used in the -e^Xpress™ platform is a breakthrough innovation that has the potential to revolutionize the production of biomolecules for the pharmaceutical and biotech industries.

​enGenes-e^Xchange™

The enGenes-e^Xchange™ technology offers a sustainable solution for antibiotic resistance marker-free recombinant protein production, using modified plasmids and E. coli strains that allow for antibiotic-free manufacturing of recombinant proteins and plasmid DNA production.

The -e^Xchange platform offers a viable alternative to traditional methods that rely on the use of antibiotics, resulting in safer and more environmentally friendly production.

enGenes-e^Xtra™

The enGenes-e^Xtra™ technology enables the extracellular production of recombinant proteins by secretion, utilizing the periplasmic space of E. coli and a leaky outer membrane to release the protein into the extracellular space. This approach allows for simplified purification processes and potentially higher yields of the desired protein.

enGenes-e^Xtra™ technology is not only a novel and efficient way for extracellular production of recombinant proteins by secretion, but it also offers cost-effectiveness through reduced downstream processing needs, making it an attractive option for clients seeking high-quality protein production.

Commercial applications

enGenes-e^Xpress™ derived technologies are well suited for fast-track antigen production and have been used to develop several different antigens already for human and veterinary medicine.

EnGenes is dedicated to the development of leading-edge Escherichia coli bioprocesses that are fully scalable. These are designed with large-scale applications in mind from the outset, ensuring solutions are tailored for industrial implementation rather than limited to small-scale, laboratory settings.

This commercially oriented approach is reflected in features, such as:

• Use of the growth decoupled T7 expression system in -e^Xpress™ as a superior alternative to growth associated production systems
• Emphasis on fermentation process development at highest-titer possible scale and yields, without need for cumbersome media optimization
• A fermentation process that can be implemented with standard fermentation equipment
• Use of host RNA polymerase inhibitor peptide
• Adaptability of the technology to various strains of Escherichia coli
• Suitability for continuous manufacturing methods
• Improved genetic stability due to lack of cell division
• Easier production of growth-interfering or toxic proteins/plasmids

Higher Yields

These technologies are fully scalable from lab to industrial quantities with 17m³ scale achieved at one client facility.

Examples of enGenes-e^Xpress™ protein and other yields include

• 13 g/L (cytoplasm) for soluble enzyme (hydrolase)
• >16 g/L (cell-free supernatant) for an affinity ligand
• >12 g/L (cell-free supernatant) for an endopeptidase
• 1 g/L (cytoplasm) for a difficult-to-express glycosyltransferase in soluble form
• >20 g/L for green fluorescent protein (GFP)

Resources

Click on enGenes expression technologies to learn more.