Recombinant Protein Production and Purification: cell line and process development

ARTES Biotechnology is one of Europe’s most innovative specialists in microbial cell line development for recombinant protein production and for production processes.

As new applications open up in research, industry, diagnosis, as well as in human and veterinary medicines, the demand for innovative solutions in the production and purification of recombinant protein has increased. Recombinant protein expression laboratories must constantly find innovative techniques to develop and produce engineered cell lines for the production of customer protein as rapidly as possible and with maximum achievable yields, high stability and safety of the product.

ARTES utilizes a range of advanced techniques to deliver production cell lines and processes with increased quality, yield and safety of customers products. The company mainly focuses on cell lines based on methylotrophic yeast Hansenula polymorpha (syn. Pichia angusta), or Escherichia coli (E. coli). Process development ensures robust and affordable manufacturing and favourably avoids the need for extensive methanol feeding, use of antibiotics and addition of animal derived components.

In order to do this, ARTES has configured its laboratories to test as many conditions for the development of cell lines and production of recombinant proteins as possible within the smallest time frame. The company has developed distinctive solutions for cost-effective production processes using both yeast-based and E.coli technologies, leveraging from cutting-edge genetic engineering, combined with a broad expertise in bioprocesses to deliver very high stability, purity and productivity.

Recombinant protein production platforms

ARTES Biotechnology operates in the field of microbial-cell line development for recombinant process development and protein production, based on powerful yeast and bacterial expression platforms.

Hansenula polymorpha (syn. Pichia angusta) methylotrophic yeast
The methylotrophic yeast system Hansenula is ARTES’ preferred technology platform and one of the leading industrial production system worldwide. For application in Food or Veterinary industry, an antibiotic resistance free host/vector system is applied. Use of animal derived components for the development of cell lines is absolute avoided and the fermentation processes are free of methanol feeding.

ARTES´ Hansenula yeast is globally applied for GRAS and Halal products, approved by international regulatory bodies such as US Food & Drug Administration and European Medicines Agency and is recommended by the World Health Organization for mass vaccination programs.

ARTES’ toolbox features a large number of expression vectors for stable, high copy integration, as well as a variety of secretion signals and chaperones to boost export of authentic and correctly folded products into the cell free culture supernatant. The Hansenula yeast expression platform is excellent in use for high yield production of intra-cellular soluble proteins as well as for the secretion of proteins. Hansenula cell lines are operated in pharmaceutical and nutritional production processes for more than 25 years at 100 % stability.

Secreted products can be harvested from cell free culture supernatants starting already virtually free from contaminating host cell proteins (< 10%), allergens, toxins and viral contaminations, thus facilitating dramatically the subsequent purification development.

Escherichia coli (E. coli) protein production
For the expression of a recombinant protein in a bacterial system, ARTES provides a highly versatile E. coli platform. The platform is characterized by an extensive host/vector toolbox for optimal expression and ability to release recombinant proteins into the periplasmic space and even into the culture supernatant.

The secretion property of the ARTES E. coli system translates directly into a higher efficiency of the production process and into reduction of costs for manufacturing of the recombinant proteins.

Process development

Expression Conditions (from gene to protein …)
Optimization of production of recombinant proteins is dependent on the number of solutions available for varying parameters. ARTES Biotechnology’s deep experience in the production of cell lines, purification of recombinant proteins and development of production processes allows it to gain from numerous technical advantages to deliver the highest chance of producing the protein of choice along with proper purity, good yield and low costs. ARTES can undertake all stages of process development to get the customer product from “gene to protein of choice”

Generation and characterization of preMaster Cell Banks
ARTES can provide preMaster Cell Banks of the production cell line selected, generated with TSE free certified materials under non GMP conditions. The preMCB comes with the full set of cell line history documents and will be tested for identity (including genomic characterization), microbial purity and productivity.

Recombinant Protein Production
The increasing worldwide requirement for recombinant protein expression, purification and process development results in unique strategies to enhance purity, yields, quality of protein production and costs of production processes.

ARTES employs many methods for development of an efficient and cost sensible production process, which are subject to the nature of the protein, the expression conditions and the future application of the protein.

Fermentation processes developed are based on synthetic media absolutely avoiding expensive additives like methanol for yeast based production or IPTG for E.coli based production and aiming at short production times (approx. 45 – 75 hrs. for yeast; approx. 24 hrs. for E. coli).

The process development at ARTES is designed right from the start to meet cGMP and large scale requirements in later-on routine manufacturing. Also here, special emphasis is given on reproducibility and cost reduction.

In-process control and final characterization of the product and of host and also process related impurities are included in all process development steps. If applicable, this characterization is performed according to international pharmacopoeia.

ARTES provides to the client the full set of documentation necessary to transfer the process to his manufacturing site or to a CMO of choice. Product samples generated and arms-length assistance in the transfer of the process developed areas well included in the tech-transfer offer.

Drawing on more than 25 years’ worth of experience in expression, cell line and process development, ARTES can provide strategic insights on technologies to be adopted corresponding to the type of protein and the clients applications.