Luina Bio’s manufacturing facility offers the latest fermentation technologies supported by world-renowned expertise in drug development and production
Microbial fermentation is the basis for the production of a wide variety of pharmaceutical drugs and biologics, from anti-cancer drugs and vaccines to hormonal disorder therapies and many others.
Over the past 10 years, more half of the 150 recombinant pharmaceutical products approved for human use by the US Food and Drug Administration and The European Medicines Agency were manufactured via microbial fermentation using bacteria or yeast[1].
The global market for protein-based biologics derived from microbial fermentation, valued at US$44 billion in 2017, is projected to grow to US$60 billion by 2020, while the market for peptide hormones and vaccines is expected to increase from US$18 to US$28 billion and US$10 to US$19 billion, respectively, over the same period[2].
Microbial fermentation in bacteria and yeast
The first protein biologics were developed by inserting the gene encoding the desired protein into Escherichia coli (E. coli), a bacterium that usually lives in the intestines of humans and the gut of some animals. Today, insulin, growth hormone, and Neupogen (G-CSF or granulocyte colony stimulating factor) are produced in this manner.
The advantage of using E. coli is that proteins can be made using a well-characterized production system that is relatively cost-effective and easy to scale-up through the fermentation of the fast-growing cells in bioreactors. Furthermore, the manufacture and purification of protein-based therapeutics have an extensive regulatory and commercial track record.
However, not all proteins can be manufactured using current E. coli production technology and misfolding of the protein molecular backbone or the inability of the host cellular systems to modify the protein background structure occurs. Adding the necessary carbohydrates and other functional groups contribute to the activity of the macromolecule, referred to as “post-translation modification.
Without these post-translational modification steps, many proteins are unable to fold into their three-dimensional shape, which determines how they interact with other proteins for biological activity. Many complex protein macromolecules, like human monoclonal antibodies, cannot be adequately produced in E. coli. and require extensive post-translational modification. They can, however, be produced in phylogenetically higher Eukaryotic organisms such as yeast, fungi and mammalian and plant cells.
For example, Saccharomyces cerevisiae (S. cerevisiae), a yeast used for millennia in winemaking, baking and brewing, has also been widely used in the manufacture of biologics. S. cerevisiae benefits from its eukaryotic model system, which enables the production and proper folding of many human proteins that would not be adequately processed by E. coli.
Furthermore, in S. cerevisiae, unlike in E. coli, the biopharmaceutical proteins can be secreted to the extracellular medium, which facilitates subsequent purification. S. cerevisiae can also perform post-translational modifications of the protein, including proteolytic processing of signal peptides, disulphide bond formation, subunit assembly, acylation and glycosylation.
One of the limitations with the use of the S. cerevisiae, however, is that it performs high-mannose type N-glycosylation and confers a short half-life of the modified protein in vivo, which produces a reduced efficacy for some therapeutic uses.
Pichia pastoris (P. pastoris) is another commonly used yeast in pharmaceutical manufacturing and is used in the production of vaccines, antibody fragments, hormones, cytokines, matrix proteins and biosimilars. A robust, durable and cost-effective yeast expression system, P. pastoris grows on simple media and secretes low amounts of endogenous protein, making it easier to recover and purify the desired recombinant protein from the cell supernatant.
Pichia has two main advantages over S. cerevisiae in laboratory and industrial settings. Firstly, Pichia is a methylotroph and can grow with the simple alcohol methanol as its only source of energy — a system that is cheap to set up and maintain — and can grow in media containing only one carbon source and one nitrogen source.
Secondly, Pichia can grow to very high cell densities, and under ideal conditions can multiply to the point where the cell suspension is practically a paste — a significant advantage when trying to produce large quantities of protein without expensive equipment.
However, several proteins require chaperonins for proper folding. Pichia is, therefore, unable to produce proteins for which the host lacks the appropriate chaperones. Additionally, Pichia has been reported to produce hyperglycosylations of several proteins, which makes it unsuitable for the production of molecules for applications in structural biology.
Luina Bio’s fermentation services
Luina Bio, an Australia based contract manufacturing organization, offers microbial fermentation services for drug development and manufacturing to the pharmaceutical, biotech and veterinary industries and has extensive experience and considerable expertise in the use of E. coli, S. cerevisiae and P. pastoris.
The company has more than 20 years of experience in the manufacture of recombinant proteins, whole-cell vaccines, viral vaccines, , and live biotherapeutics for use in pre-clinical, phase 1 to 3 human clinical trials for customers worldwide . We also producecommercial veterinary products for clients within Australia and internationally.
Luina Bio’s manufacturing facility in Brisbane utilizes TGA/APVMA licensed processes and technologies operating to international Current Good Manufacturing Practices (cGMP) standards that provide comprehensive solutions for both biological and small molecule drugs
The facility houses fermentation vessels ranging from 5L to 500L that can be operated in numerous modes, including batch, fed-batch and perfusion and offer a full suite of manufacturing for both whole-cell preparations, recombinant proteins from microbial hosts and live biotheraputics (both aerobic and strictly anaerobic BSL1 and BSL2).
By adopting cGMP fermentation procedures, Luina Bio builds quality into every stage of the manufacturing process, ensuring that regulatory requirements are met in terms of safety, product identity, quality and purity, and can provide batches of cGMP material produced in a range of microbial expression systems.
The company offers world-leading fermentation and related services in:
- Master and working cell bank preparation, validation and storage
- Fermentation and subsequent downstream purification
- Up and downscale fermentation optimization; and
- Downstream processing optimisation and development
[1] Rios, Maribel. “A Decade of Microbial Fermentation.” Bioprocess International (2012).
[2] Dewan, Shalini Shahani. “Global Markets and Manufacturing Technologies for Protein Drugs.” BCC Research (2016).
