These enGenes microbial expression solutions are based on its unique and proprietary enGenes X-Press bacterial cell line technology platform, enhanced by patented manufacturing processes to leverage the basic X-Press growth decoupled advantage. The net results are substantially higher soluble product yields and easier downstream processing (DSP), resulting in up to 80% reduction in cost of goods sold (COGS).
Biotech manufacturing challenges
Bioprocesses, harnessing the properties of cells and enzymes through tools such as recombinant DNA to develop novel products, presents a series of challenges, particularly in upstream and downstream processing as well as scale up to industrial viability.
Manufacture of a typical recombinant DNA-derived product will involve up to 20 steps between identifying basic source materials or tissue and final packaging. The initial ten or so steps are concerned with genetic manipulation of the host organism; such as through cloning a gene from DNA into Escherichia coli (E. coli). This will involve genetic engineering at laboratory scale to establish levels of expression available and the stability of the constructed strains, defined as growth and production characteristics of the recombinant cells as a function of the culture environment, with parameters such as cell growth rate, specific productivity, and product yield calculated to measure the performance of the organism.
Once viable culture conditions for production are established, process development will involve using a benchtop bioreactor to establish the ideal inducer, induction strength, temperature, pH, dissolved oxygen concentration, stirrer speed, and other process variables. The challenge here is that moving the cell culture to a stirred bioreactor may not speed the process but harm it. The viability of the process will be defined in measured and calculated parameters such as mass transfer coefficients, mixing time, gas hold-up, oxygen uptake rate, power number, energy dissipation rate, etc. It must also be decided whether the culture is best operated as a batch, semi-batch, or continuous process.
The production strain must then be suitable for tech transfer to CDMO or CRO pilot scale (100- 1,000 litre up to m3-scale) bioreactor, where equipment design or configuration may affect process viability. Full scale contract manufacturing also needs to consider downstream process (DSP) product recovery, which can be a major source of added cost and lost efficiency. For some recombinant-DNA-derived products, DSP purification can account for 80 to 90% of the total processing cost.
The unique enGenes-X-press™ proprietary technology platform for outstanding recombinant protein production in Escherichia coli (E. coli) uses an advanced growth decoupled mechanism platform that allows customers to achieve cost-effective production of recombinant proteins in microbial expression systems.
The technology is leveraged from enGenes’ world-class experience in cutting-edge synthetic biology and genetic engineering, combined with a broad expertise in bioprocess engineering, to produce a range of best-in-class protein expression systems.
The X-press technology platform is based on a genetically modified host cell based on E. coli B-strain linage (BL21), and recently also K-12 strain derivates, that allow for introduction of standard expression vectors (with T7 promoter) with no need for cumbersome adaptations. The developed fermentation process can be implemented with standard 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. This enables the production of difficult-to-express proteins.
The technology 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. It also enables recombinant product formation to be decoupled from cell growth, enabling bioprocessing with clear separation of biomass growth and product formation. This in turn makes it possible to generate significantly higher specific and volumetric yields compared to the previous state-of-the-art (Escherichia coli BL21 (DE3).
X-Press uses a two phase production process with accumulation of biomass, followed by production of recombinant protein. Along with high specific yields, this process offers improved scalability and controllability, robustness, fewer optimization steps and shorter development cycles across the whole pharmaceutical contract manufacturing process.
X-Press is a versatile core technology that can be intelligently applied in many ways in conjunction with other enGenes products to produce proteins, enzymes, bioconjugates, metabolites, small molecules, and plasmid DNA (pDNA) or RNA.
These related technologies include:
- -X-tra for extracellular secretion of recombinant proteins
- -X-pand for expansion of genetic code and incorporation of NCAAS
- -X-cite for continuous production of biomolecules
- -X-cess for pDNA upstream and downstream processing and growth-decoupled pDNA production
- -X-change for antibiotic resistance market free protein of biomolecules.
Used in combination, this suite of technologies has potential to increase yields up to five-fold or beyond.
Contract development & manufacturing advantages
enGenes operates in the role of a contract research organisation (CRO) that is able to partner with chosen contract manufacturing organisation (CMO) or contract development and manufacturing organization (CDMO).
Involving enGenes at the initial research and development phase has clear advantages over using conventional CDMO pharma services, which tend to use standard processes that are not closely aligned to the characteristics of the target protein or molecule. Nor are pharmaceutical contract manufacturing CDMO or CMO companies well-motivated to achieve high yields, since these will mean fewer manufacturing campaigns, with loss of revenues.
In contrast, enGenes possesses the technologies, skills, training and motivation needed to develop the most cost-effective process that is ready for out-licensing or tech transfer to a full form CDMO or always having in mind that it has to be practicable at a larger contract manufacturing organization (CMO).
Even in advance of the project research phase, enGenes is able to liaise with customers chosen CDMO or CMO manufacturing organisation to ensure that the developed process exactly fits with the capabilities of the CMO. It will ensure that the tech transfer process runs smoothly with upstream and downstream pre-developed and precise yield estimates for fed-batch processing so that client gets the best value for money.
The end result is to deliver the best manufacturing process available with huge potential savings overall for pharma, industrial biotech or alt-protein companies.
For clients who wish to produce products in-house, enGenes can provide detailed advice on what equipment to use for each particular task so that development can move smoothly into production.