Robust and scalable fermentation & protein purification
Pichia protein production processes at VALIDOGEN show extraordinarily high space-time yields. Multi-gram-per-liter titers are often reached in the culture supernatant (depending on the protein of interest, up to a record of 35 g/L) prior to downstream processing.
Starting from validated VALIDOGEN protocols our Pichia cultivation experts can develop an optimized production process for your protein. We take into account scale-up considerations to elucidate key parameters and deliver a robust and scalable process. Our customers have successfully scaled VALIDOGEN protein production processes to up to 100,000 L cultivation volume.
VALIDOGEN can also provide full protein purification process development. In close coordination with our customers we will evaluate chromatography steps, determine capacities, maximize yield and purity, and establish clearance of impurities.
VALIDOGEN’s 15+ years of experience have led to the development of robust cultivation protocols allowing Pichia strains with UNLOCK PICHIA® technology to maximize their productivity potential and reduce cost of protein manufacture. Starting from extensively tested and high-performing in-house protocols we can further adapt parameters to be optimal for your protein and application.
In our suite of parallel bioreactors and microbioreactor system we can undertake development and optimization work for your process with short lead times. From small process parameter modifications to extensive Design of Experiment investigations a process customised for your protein will be developed, always with manufacturability in mind.
In addition to our highly efficient methanol-induced promoter variants and sophisticated fermentation protocols, we develop effective methanol-free processes based on VALIDOGEN's methanol-free promoter variants. Methanol-free cultivation offers advantages of avoidance of hazardous chemicals, more economical production plant requirements, no metabolic burden from change of carbon source, and reduced aeration and cooling demand. VALIDOGEN have established methanol-free protein production strategies to shorten process time (~60h) which allows for the maximization of space-time yields.
To allow our customers to quickly receive a small sample of their protein for evaluation we can undertake rapid set-up of a purification process. In a short time we can provide highly pure milligram to gram amounts of your protein from a Pichia culture supernatant. Considering the properties of the protein and our experience we will scout multiple methods to identify the right purification strategy to achieve the desired specifications. Purification methods often include standard chromatography techniques such as ion exchange or hydrophobic interaction as well as affinity chromatography, and may be combined with ultrafiltration steps.
Our DSP team can also take downstream processing one step further by providing full, in-depth process development for a robust, scalable process. In close coordination with our customers we will evaluate capacities of individual chromatography resins, maximize yield and purity, establish clearance of impurities, and understand how process steps are interlinked. Comprehensive documentation and support guarantee smooth transfer of the optimized downstream process to your production site.
All DSP activities are accompanied by analytical characterization of process intermediates and purified product. Established in-house methods include PAGE, capillary electrophoresis, HPLC, Western Blot, and ELISA, as well as methods to quantify process-related impurities including HCP and host cell DNA.
Early development enables you to optimize strain performance and fermentation conditions before moving into scale up, or expensive GMP environments. This approach reduces risk, and ensures a smoother, more cost-efficient transition into manufacturing.
Yes. Even if the strain was developed elsewhere, we can apply our expertise in Pichia fermentation to optimize expression levels, productivity, and process robustness. This often results in significantly higher yields and lower costs compared to unoptimized processes.
Yes. Our development is always performed with scalability in mind. Processes optimized in our labs are designed to transfer smoothly to industrial fermentation facilities, maintaining productivity and product quality at large scale.
We develop processes at scales up to 10 L bioreactors. VALIDOGEN is not a GMP facility — instead, we deliver tech-transfer-ready processes that can be handed over to your selected CDMO for GMP manufacturing. We have undertaken many such transfers.
A tech-transfer-ready process is one that has been well characterized and optimized, with robust fermentation conditions, reproducible performance, and thorough documentation. This ensures that a CDMO can implement the process at larger scale without additional major development steps.
We apply a broad range of chromatographic and non-chromatographic purification methods depending on the target protein and its application. Chromatographic techniques typically include ion exchange, hydrophobic interaction, mixed mode, and affinity chromatography, often combined in multi-step strategies to achieve the desired purity and yield.
In addition to chromatography, we can apply diafiltration, ultrafiltration, precipitation, and extraction methods as part of the purification workflow. These approaches can simplify the process, reduce costs, or serve as preparatory steps before final chromatographic purification.
Purity levels depend on the protein and intended use. In most cases, our workflows achieve high-purity protein suitable for research and preclinical applications. For industrial enzymes, purification can be tailored to cost-effectiveness rather than maximum purity, while for therapeutic candidates, very high purity levels can be reached through optimized multi-step processes.
VALIDOGEN's expert team is happy to discuss your project and help find the right solution. Send any project related requests to project@validogen.com, directly to our scientists, or use our contact form.
