Given that several major contract development and manufacturing organizations (CDMOs) such as Catalent, Patheon, and Samsung Biologics don’t even offer protein production in microbial cells, positioning yourself purely as a microbial CDMO for producing protein-based biotherapeutics might seem a little unconventional.
However, according to Gerhard Schmid, PhD, president of Wacker Biosolutions, in a 2017 HTB Biopharmaceutical Contract Manufacturing report the microbial CDMO market was estimated to be growing by 9% per year and is projected to reach $1.6 billion dollars by 2022. With the total recombinant CDMO market of mammalian and microbial cell culture projected to be $6 billion by 2022, then concentrating on a 26% slice of that pie might not sound such a bad idea and Wacker could be on to something with their niche market approach.
Wacker Biosolutions, Wacker Group’s biotech and fine chemicals division, recently announced 2018 sales of 227 million Euros; 33% of these sales were due to pharma, mostly biopharmaceuticals. According to Schmid, since 2014, the business line’s sales have also grown by an average of 19% per year. In fact, the division did rather well when compared to other Wacker traditional business divisions such as Wacker Polysilicon, where global competition and raw materials costs are having a major impact on profits. “Wacker Biosolutions may be one of the the smallest of the Wacker divisions but it is one of the most exciting,” Schmid says.
Wacker Biosolution’s biopharmaceuticals business line specializes in the manufacture of three main product types: recombinant protein therapeutics, live microbial products (LMPs), and vaccines. The firm’s expertise in microbial production of biopharmaceuticals was acquired using a buy-and-build strategy. This began in 2005 when it acquired Prothera and then Scil Proteins in 2014. Both plants are run by the group’s subsidiary, Wacker Biotech GmbH headquartered in Jena, Germany. The latest production site purchased was Synco Bio Partners, which was acquired and renamed Wacker Biotech B.V. in 2018. The three sites combined offer full-service contract manufacture from cell banking and process development, fermentation, and purification in both traditional stainless steel and single-use equipment through to fill-finish in vials.
The majority of therapeutic products manufactured by these CDMOs are recombinant proteins and Susanne Leonhartsberger, PhD, vp, business line biopharmaceuticals at Wacker Biosolutions, cites AGC Biologics, Boehringer Ingelheim, Fujifilm Diosynth, and Lonza as their main competitors for therapeutic protein production. Table 1 shows the range of microbial cells where each CDMO specializes and, unsurprisingly, E. coli and P. pastoris are the most popular microbial cells on offer.
The largest group of therapeutic proteins are monoclonal antibodies (mAbs), with the market for these valued at around $100 billion in 2018. However, as Schmid explains, “Most mAbs are glycosylated and therefore cannot be produced in microbial culture, so we can’t compete in traditional mAb production.” Currently, of the over 70 mAbs that have been approved for clinical use, only two commercially available antibody fragment (Fab) products, Lucentis (ranibizumab) to treat age-related macular degeneration (AMD) and Cimzia (cerolizumab pegol) to treat autoimmune diseases, including rheumatoid arthritis, are manufactured in E. coli.
Lucentis biosimilars are being developed by thirteen companies globally using E. coli for the manufacturing and these include front-runners Formycon, Pfenex, and Samsung Bioepis and smaller biotechs such as Mabion. Wacker Biotech is active in this area and is in collaboration with a large undisclosed Asian biopharma company.
Super charged E. coli
Wacker Biotech offers two patented technologies for production of therapeutic proteins using E. coli, and these are known as ESETEC® and FOLDTEC®. ESETEC is based on an E. coli K12 strain which has been genetically engineered to secrete protein directly into the culture medium instead of keeping the proteins in the periplasm as most traditional E. coli strains would.
According to Guido Seidel, PhD, managing director of operations at Wacker Biotech, “Because ESETEC allows direct secretion of proteins, it reduces the number of processing steps. For example, there is no need to homogenise or sonicate cells and remove cell debris to get the protein in solution. Since there are fewer process steps this results in a higher yield. We see yields typically of 2 g/L from 200 L fermenters for Fabs and up to 16 g/L for other protein classes using ESETEC and this is around four times higher than many E. coli strains produce. The advantage of this is it dramatically reduces the cost of goods (COGs) to produce a protein therapy.”
The types of protein therapeutic commonly being produced using ESETEC include Fabs, and Seidel presented a case study which showed that ESETEC has been used from scratch to GMP manufacturing in 1500 L in collaboration with MedImmune to manufacture a Fab fragment which reverses the effect of a drug, anti-platlet antidote that binds to Ticagrelor.
“MedImmune is so pleased with the results that it licensed ESETEC, which improves yields and lowers COGs, for producing this Fab,” says Seidel.
The FOLDTEC technology is also based on E. coli and produces proteins that have the correct 3D structure for binding and therapeutic activity. The technology uses E. coli producer strains which can be cultured without any antibiotics and feature phage-free plasmid maintenance combined with proprietary process refolding conditions (including specific temperatures, buffers, and pH). An example of a marketed protein therapeutic being produced using FOLDTEC technology is Reteplase, a recombinant form of a tissue plasminogen activator protein used to treat myocardial infarction.
Vaccines and LMPs
As well as manufacturing recombinant proteins, through the acquisition of its Amsterdam plant, Wacker has extended its capabilities and now manufactures LMPs and vaccines using microbial fermentation. The vaccine market (valued at around $33.70 billion in 2018) is worth considerably less than the protein therapeutics market and is also biased towards the use of mammalian cells lines such as Vero for production. However, there is still work it would seem for CDMOs producing vaccines using microbes.
The CDMO currently works with 10 different organisms including pathogenic bacteria: V. cholera, Salmonella typhi, and Listeria monocytogenes, as well as probiotics including Lactobacillus spp. Jörg Lindemann, PhD, managing director of Wacker Biotech B.V. says, “The Amsterdam facility was originally built for API production. Later, vaccine production was added. For example, PaxVax, a vaccine to prevent cholera, is manufactured here using live, attenuated bacteria. The vaccine gained FDA approval in 2016 showing that we can manufacture to high standards.”
He adds, “We have 270 L and 1500 L stainless steel fermenters, as well as 20 L to 200 L Wave bioreactors and 250 L single-use bioreactors, which we use for the production of LMPs. These are a promising new class of biologics, which can be used as vaccines or as therapies. On the therapy side, we are manufacturing a bacteria that could improve wound healing for diabetics, and this is due to enter clinical trials in 2019.”
Much of the process development and cell count work being performed at the Amsterdam site is manual as many of the large scale fermentations are operated in standard conditions at pH 7. Therefore, process development work is minimal and is carried out in traditional shake flasks, small-scale fermenters and colony counting is also being done by semi-automated plate counts. According to Lindemann, automation including the use of single-use bioprocess mimics is an area that Wacker Biotech B.V. is looking to invest in.
“When it comes to developing biotech processes, we have more than 200 sucessfully completed projects. We work with many leading biopharma companies including Chiesi, Novartis, Roche, MedImmune, and UCB and can manufacture to GMP standards. We are now the biggest dedicated microbial CDMO in the world and will continue looking at expansion possibilities where possible, as we want to be up there with the other major CDMOs,” Leonhartsberger says.
Schmid concludes: “The investments and acquisitions of the past months present a huge opportunity because we can now show that we are a chemical company capable of operating a large part of our business on a biotechnological basis. We have the ability to take on new customers, as our production facility in Amsterdam doubles our capacity. We are still looking at mammalian cells production as it is a more lucrative market than microbial but because our expertise is in microbial, I think unless a CDMO of the right size with good mammalian culture capabilities becomes available, it is likely we will remain a dedicated microbial CDMO.”