We understand that a successful biomanufacturing process development project not only requires Upstream Processes (USP) but also Downstream Process development (DSP).
The Downstream process in a biotechnology fermentation process comprises all stages from harvest and primary capture through to the final product.
A downstream process workflow can include:
- Cell separation - Centrifugation, Membrane filtration
- Cell disruption - Homogenisation, Sonication
- Extraction - Solvent extraction
- Isolation - Enrichment of the target product
- Purification - Column chromatography
- Drying - Lyophilisation or spray drying
Our approach
We adopt stage-appropriate approaches to downstream process development. We tailor our work to best fit your current operational scale and future aims.
DSP approaches:
- Rapid contract DSP: A quick and effective lab-scale DSP to enable the supply of compound as soon as possible.
- In-depth DSP development: A fully developed DSP workflow that is ready for tech transfer to a CDMO.
We understand the importance of establishing a robust and functional downstream process (DSP) well before scale-up. When launching a downstream process development work package, understanding your priorities and current operational scale is key.
Client requirements often include:
- Maximising yield
- Meeting specific quality criteria
- Cost reduction
- Product supply
Process scalability is key
We keep scalability and potential commercial rollout in mind throughout the downstream development process. From product capture and primary extraction through to final product purification, ensuring industrial viability is imperative.
Downstream process development methods
We have a range of DSP methods at Isomerase. Our experienced team will carefully evaluate your requirements and select the most effective techniques to ensure optimal results.
Examples of our downstream process development services include:
- Homogenisation (various scales)
- Periplasmic preparation
- Extractions (solid phase, liquid/liquid)
- Flash chromatography and High Pressure Liquid Chromatography (HPLC), normal and reversed-phase
- Liquid Chromatography + Mass Spectrometry (LC-MS)
- Size exclusion chromatography (SEC)
- Ion-exchange chromatography (IEX)
- Affinity chromatography (AC)
- Hydrophobic interaction chromatography (HIC)
- Resin capture
- Precipitation
- Crystallisation
- Microfiltration
- Ultrafiltration
Downstream process development projects
Examples of DSP projects we have carried out include:
- Scalable DSP Development - We design and optimise robust downstream processes (DSP) that are scalable and ready for tech transfer to CDMOs.
- Material Isolation & Supply for Testing - Our isolation capabilities, from milligram to gram scale, have been utilised across multiple previous projects. Once isolated, these compounds (in solution or powder form) can be shipped directly to you for any further analysis required.
- Isolation of Trace-Level Compounds - Our chemists are highly experienced at isolating compounds from complex mixtures, using techniques such as preparative HPLC. Furthermore, we can isolate low-level background contaminants 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 - While not a core capability, we provide in-house synthetic chemistry support and manage external chemistry teams to meet project needs.
- Cell Bank Production - We produce and qualify Master Cell Banks (MCBs) and Working Cell Banks (WCBs)
- Product capture - Our teams can optimise and develop scalable primary product capture and extraction methods
- In-process controls - Determination and validation of robust and predictable in-process control points.
- Hold Point Analysis - We conduct stability studies on intermediates to determine optimal storage points within the DSP workflow.
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, a bacterium, and Pichia, a yeast.
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 involves the production of a target compound at an intermediate scale. This scale provides a unique opportunity to bridge the gap between small lab-scale (3-7 litres) vessels and commercial production-scale vessels. At Isomerase, we frequently use pilot-scale (20-30 litre) vessels to validate the fermentation process’s scalability while monitoring fermentation parameters for further optimisation. We can fine-tune various parameters such as medium composition, fermentation conditions, and feeding strategies.
It is vitally important to optimise and validate processes at pilot scale to derisk commercial production runs. Production runs at commercial scale require a very large investment, as all costs scale with the fermentation volume. Before making such a large investment, a process must be fully understood and robust.
When scaling up to commercial production vessels, mixing characteristics are a key focus. The agitation motor's power and the shape of the vessel itself often change as fermentation vessels grow. To ensure aeration is consistent, the distribution of bubbles within the broth is important. Without maintaining this crucial growth condition, we cannot be confident that growth rates and, by extension, production titers will scale appropriately.
Our pilot vessels mirror the aspect ratios of commercial tanks. We often scale processes from ~2 litres to ~20 litres, fermentation volume, using metrics that can be easily transferred to larger vessels (mixing impeller tip speed).
Fermenting at pilot-scale is further beneficial for:
- Testing downstream processes at a larger scale
- Increased material supply for our clients
- Additional product testing and analysis. This may include product stability studies, bioactivity assessments, and formulation optimisation
- Producing sufficient product quantities for market testing and validation
In summary, pilot-scale fermentation serves as a crucial step in the development and commercialisation of fermentation-based products as it helps optimise the process, validate scalability, and gather market feedback.
Why choose Isomerase for downstream process development?
At Isomerase, we specialise in both upstream (USP) and downstream (DSP) process development, from lab-scale microbial fermentation to pilot scale.
What sets us apart is our collaborative approach. Our DSP team includes molecular biologists, bioprocess engineers, and analytical chemists, bringing together a wide range of expertise. This cross-functional collaboration starts at the proposal stage and continues throughout the project lifecycle, ensuring that every process we develop is technically sound and economically feasible.
This integrated approach allows us to manage complex, multidisciplinary projects effectively and deliver innovative, biobased solutions that meet evolving industry demands