We understand that a successful biomanufacturing process development project not only needs optimised Upstream Processes (USP) but that Downstream Process (DSP) development is equally important.
The Downstream process in a biotechnology fermentation process comprises all stages from harvest and primary capture through to final product. This can include cell separation (e.g. centrifugation, membrane filtration), cell disruption (e.g. homogenisation or sonication), extraction (e.g. solvent extraction), isolation (generally covers initial enrichment of the target product), purification (generally covers final purification of the product to the level required for release) and drying (such as lyophilisation or spray drying).
We appreciate the importance of developing a functional, robust DSP process well in advance of transferring to scale. The first consideration in DSP development is understanding the client’s requirements. That may be maximising yield, specific quality criteria or a cost of goods barrier.
We adopt stage-appropriate approaches. Either a quick and effective lab-scale DSP to enable supply of compound as soon as possible, or a fully developed DSP that is ready for tech transfer to a CDMO. We keep scalability and potential commercial rollout in mind for all steps, from product capture and primary extract to final purification. A key to good DSP is a full understanding of the characteristics of the product, and how it behaves in different matrices. We routinely assess the stability and solubility of the desired target at different stages of DSP, including hold point analysis and IPC strategies. For example, hold-point analysis allows us to identify stages where a product is stable before final purification, and crucially where it is sensitive to storage or delays. Other key attributes that we usually recommend assessing include activity, toxicity, size, purity, 3-dimensional structure, related compounds and contaminant profile.
We develop our processes through a combination of experience, know-how and Design of Experiment (DoE) approaches.
Examples of DSP methods that we carry out at Isomerase
We have a range of DSP methods that can be applied, depending on the properties of the product.
Examples of our DSP bioprocess development services include:
- Homogenisation (various scales)
- Periplasmic preparation
- Extractions (solid phase, liquid/liquid)
- Flash and HPLC chromatography (normal and reversed-phase)
- Size exclusion chromatography (SEC)
- Ion-exchange chromatography (IEX)
- Affinity chromatography (AC)
- Hydrophobic interaction chromatography
- Resin capture
Examples of DSP projects
Examples of DSP projects we have carried out include:
- Development of a robust scalable DSP process - a fully developed DSP that is ready for tech transfer to a CDMO
- Isolation and supply of material for testing - we have isolated compounds and other materials in quantities from milligrams to grams in-house on a wide variety of projects. These have been shipped directly to partners or we have helped manage screening cascades by storing materials at Isomerase and sending them directly for biological or chemical testing.
- Isolation of low-level background products - our chemists are highly experienced at isolating compounds from complex mixtures, using techniques such as preparative HPLC. We have supported partners by isolating low-level background contaminants, for example, to support regulatory filings.
- Semisynthetic chemistry on Natural Products - we have supported drug discovery analogue campaigns by carrying out semisynthetic chemistry in tandem with the generation of bioengineered analogues.
- Chemical synthesis of small molecules - whilst it is not a core capability, we are able to carry out in-house synthetic chemistry to support projects and have managed external chemistry teams on behalf of partners.
- Cell banks - we can produce and qualify Master Cell Banks (MCBs) and Working Cell Banks (WCBs) for manufacturing campaigns
- Product capture - we can optimise and develop scalable primary product capture and extraction
- In-process controls - Determination and validation of robust and predictable in-process control points
- Hold point analysis – Stability studies on intermediates throughout a DSP process support assessment of the best points in a process to store material safely and reliably.
The Biomanufacturing Process
Biomanufacturing involves engineering a cell to produce a specific protein. Using well-established techniques to transfer a gene encoding the desired protein into a ‘production cell’. One of the most commonly used microorganisms is E. coli bacterial cells though the yeast Pichia is increasing in use.
One of the first steps in the biomanufacturing process is selecting or developing a suitable microbial strain or cell line that possesses the desired characteristics to produce the target product. This can involve genetic engineering techniques to optimise the organism's metabolic pathways or introduce specific genes for increased production – Isomerase can also assist you with this.
Once the strain is selected, banked and qualified, it is grown in a laboratory-scale seed culture to produce a sufficient quantity of biomass. This seed culture is then transferred when grown to specified parameters to a production vessel, such as a bioreactor, where the cells are grown under controlled conditions to maximise product formation.
What are the main uses of pilot-scale fermentation?
Pilot-scale fermentation refers to the production of a product using fermentation techniques on a small scale, typically larger than laboratory-scale but smaller than full-scale commercial production.
Pilot-scale fermentation allows engineers to optimise and refine the fermentation process before companies need to invest in large-scale production. It provides an opportunity to test and fine-tune various parameters such as medium composition, fermentation conditions and feeding strategies.
It can also provide an opportunity to produce larger quantities of their target product for further development and characterization. It allows for more extensive testing and analysis, including product stability studies, bioactivity assessments, and formulation optimization.
Before launching a product on a commercial scale, businesses may want to conduct market testing and validation. Pilot-scale fermentation allows them to produce sufficient quantities of the product for testing and evaluation by potential customers or partners.
This feedback helps in understanding market demand, gathering user preferences, and validating the products’ commercial viability.
In summary, pilot-scale fermentation serves as a crucial step in the development and commercialization of fermentation-based products as it helps optimise the process, validate scalability and gather market feedback.
Why Choose Isomerase?
At Isomerase, we have expertise in microbial fermentation at the lab scale, pilot scale and supporting through to scaled manufacturing. We can generate and develop Upstream (USP) and Downstream (DSP) aspects of a process for a wide range of products. We know what makes a successful commercial process and include this thinking at all steps of USP and DSP development. We are committed to sustainability and use renewable resources wherever possible in our processes.
Contact us for more information about pilot scale fermentation services or any of our other biomanufacturing processes and request a quote from us today.