E. coli is a tried and true expression platform for biologics. E. coli is usually chosen based on productivity. You can potentially get up to 20g/L , although ~8 g/L is a more typical titer.
In our experience, E. coli is also preferred for some classes of molecules that could interact with mammalian expression systems. These include growth factors, cytokines and interferons.
- Fast processing. Plant time is short even if you are accounting for recovery steps. This can have an impact on cost but also minimizes the contact time with manufacturing systems.
- Flexibility. Microbial systems allow for processing with natural decoupling steps (cell paste for intracellular soluble and inclusion bodies). This increases the opportunities to assign manufacturing slots in a multi-product facility. It also allows for staggering of readiness activities which can decrease time to manufacture.
- Materials can be more cost effective and are less blocked by proprietary issues, (i.e., cell culture feeds). Most formulations are made from individual components that are controlled and understood.
What are particular challenges of working with E. coli?
E. coli post-translational modifications are limited if non-existent. Another one of the challenges with E. coli lies with getting product out of the cell. In our experience we have found that we may have have to homogenise to break open the cell (i.e., typical pressures ~800 bar). This has the disadvantage as it’s another processing step, it makes a “soup of protein” to clean up, puts heat into the culture, and may release proteases that might degrade the therapeutic protein.
If we express as inclusion bodies however, then homogenization and IB washing is a very quick and efficient way to get 90% plus purity in a simple step. Clearly however, the disadvantage with this is we need a refold post this step. At FUJIFILM Diosynth we have extensive experience and solutions to address these challenges.
What do we bring to your E. coli program?
One of our biggest differentiators as an E. coli CDMO is our pAVEway™ expression platform. It is a tried and tested expression platform with over 90+ pAVEway studies completed. We have achieved significant titers in fully defined growth media, therefore robust and consistent. We also offer our customers with a well developed fermentation system via the ambr250, Caliper, and TECAN. This system has been proven to scale up to our cGMP Manufacturing facilities. In some instances, E. coli manufacturing could have larger barrier-to-entry with substantial utility and equipment requirements. As a CDMO we are a fully stocked microbial outfit with decades of experience with microbial manufacturing of recombinant products. This positions us nicely for microbial programs.
How do we help?
With extensive E.coli development and manufacturing experience and a team of expert scientists, FUJIFILM Diosynth Biotechnologies offers a full range of services including: strain development using our pAVEway™ expression platform, process and analytical development services, combined with flexible cGMP manufacturing assets, to support you through the full life cycle of your product, from development to clinical and commercial manufacture.
We also bring extensive regulatory experience as we are licensed to manufacture two commercial E. coli expressed products.
Please find some useful references for the expression of recombinant proteins in E. coli here: