Your research is only valuable if your data can be reproduced. It is therefore crucial that high quality, well-characterized research materials are used for your preclinical studies. Our capabilities and experience absolutely qualify us to be your research partner and deliver your protein-based products.
We have significantly invested in optimizing transient transfection protocols for both HEK293 and CHO mammalian cell lines for protein production. As such, optimized protocols are available using either chemical, lipid-based, or electroporation-based methods. For the rapid delivery of gram-scale proteins, we perform high cell density, high flow electroporation (MaxCyte technology). With this method, we have demonstrated that more than 90% of the cells are transfected as demonstrated by GFP expression and, equally important, cell recovery is over 90%.
Using these protocols, we have run transfections of up to 50 L scale and delivered many high quality and high yield research batches including antibodies, growth hormones, antigens, enzymes, and interleukins. In addition, the optimized transfection protocols are also deployed to perform high-throughput small-scale protein productions to yield 10 μg to 1 mg batches. Hereto, the optimized transfection protocols are coupled to the SCOUT® technology providing the capability to generate hundreds of purified protein variants within a 4-week time frame. In such high-throughput production experiments, each protein is delivered with its own certificate of analysis containing pivotal data on crucial process parameters.
For our mammalian production platform, we deploy a wide variety of cell culture formats. For example, our capabilities include adherent, microcarrier and suspension culture. We can deploy a variety of mammalian cell lines, including CHO and HEK293. Production scales can range anywhere from small (2-20 mL) to intermediate (0.5-200 L).
From a wide variety of equipment, we can select the most suited to produce your molecule of interest. For suspension cultures it could be shake flasks, roller bottles, WAVE Bioreactor™ systems or stirred tank bioreactors, while for adherent cell cultures it could be cell factories, iCELLis® or scale-X™ bioreactors. For small-scale productions (microgram to milligram range), we typically use our SCOUT® technology. This system integrates high-throughput cell culture with high-throughput purification and matching analytical capabilities.
When larger quantities are required, we can deploy our optimized high cell density flow electroporation technology for transient productions of up to 100 L scale using our in-house animal component-free, or customer-developed medium, using either batch, fed-batch or perfusion protocols. Alternatively, we can use our STEP® technology to develop stable producer cell lines in record time or help you to further develop your manufacturing process and select the optimal growth media.
Matching our upstream capabilities, we can deploy a variety of equipment for clarification, chromatography and filtration. For example, in the purification of proteins and antibodies, we offer clarification by depth and membrane filtration. Prior to purification, concentration and buffer exchange is typically performed using flat screen tangential flow filtration. Purification is normally achieved using several chromatography steps. The chromatography methodology used is dependent on the molecule of interest with a variety of properties being exploited via affinity, pseudo affinity, ion-exchange, hydrophobic interaction, size-exclusion or mixed mode to achieve the desired purity. If required, viral clearance filtration can be performed.
Formulation is typically performed using desalting chromatography or buffer exchange using flat screen tangential flow filtration. Sterile filtration is performed using 0.2 µm or 0.22 µm filters. Finally, for analysis of integrity, purity, yield and identity of research preparations, we use our developed product-specific assays; SDS-PAGE, western blot, ELISA and various chromatography methods.