Sue Pearson Ph.D. Freelance Writer GEN

Speakers at bioProcessUK say the United Kingdom must continue to build on excellence if it is to remain a prime bioprocessing location.

BioProcessUK’s 10th annual meeting in London proved to be a time of reflection on what the U.K. is doing right and the direction it needs to head in the coming decade if it is to stay a leading center for biomanufacturing. Many speakers cited news in 2013 of Fujifilm Diosynth Biotechnologies opening a new mammalian cell culture biomanufacturing plant in Billingham utilizing single-use technologies, AstraZeneca investing £330 million ($539.4 million) to locate its research including its nonvaccines biologics activities such as bioprocess development at the Cambridge Biomedical Campus, as well as rumours that GSK is going to increase biomanufacturing capacity there1 as positive signs that the U.K. remains a top location for production of biologicals.

Mark Bustard, Ph.D., technical director of bioProcessUK at the HealthTech and Medicines Knowledge Transfer Network (KTN), a U.K. government initiative, comments: “In the past 10 years, the U.K. has become a biomanufacturing cluster similar to Research Triangle Park or Boston in the U.S. with 260 companies and around 12 thousand people actively involved in developing or manufacturing biologicals or the wider supply chain. Bioprocessing is now an industry in its own right, which has more recently become a priority area supported by the U.K. government; this makes it a good location to do the research and translate that research into products.”

Many at the conference cited good links between academia and industry as key to why biomanufacturing is so productive in the U.K. Head of the University College London (UCL) Biochemical Engineering department Professor Nigel Titchener-Hooker, Ph.D., states: “In the U.K., bioprocessing is one area where academic/industrial collaboration works particularly well and is helping accelerate life science research to commercial discoveries in regenerative medicine, vaccines, and biological drugs.” He revealed that UCL had played a pivotal role in this and that in recognition of its excellence, UCL’s Biochemical Engineering Department had recently been awarded the 2012–2014 Queen’s Anniversary Prize for Higher and Further Education, which is the U.K.’s highest national honor for innovative research.

Dr. Titchener-Hooker adds: “The cost of goods of biologics is still relatively high. We have a huge way to go and we have to find innovative ways of reducing this to ensure these types of biological therapies and vaccines are more affordable so that more of the world’s population can benefit. Therefore, continued dialogue between academia and industry is important for providing solutions to make bioprocessing faster and cheaper.”

Optimizing Clone Selection and Cell Culture Processes

One method of producing more affordable biologics discussed at bioProcessUK is to increase productivity and yields by improving upstream processing. Kripa Ram, Ph.D., vice president, bioprocess and manufacturing sciences at MedImmune explains: “In upstream processing we have worked on improving cell-line development and optimizing feed and media formulations to promote better yields. Previously, we used static multi-well plates and shake flasks to screen clones, and put a small number of clones in a benchtop bioreactors and hoped one worked. This approach was low throughput and had poor predictability as we didn’t know until we put our carefully selected clones in the scale-up bioreactor if they would perform as we expected. We knew we had to automate and improve these areas of the process to see an improvement.”

According to Dr. Ram, since swapping the static multi-well plate approach with the Life Technologies’ ClonePix FL to select clones and the Hamilton Star system for shaking deep well plates there has been a significant improvement in titer predictability. Cell culture predictability has been further enhanced by, replacing the shake flask method with the ambr microbioreactor system from TAP Biosystems, with better control of pH, dissolved oxygen (DO) and temperature, producing similar titer results to a 12,000 L bioreactor.

Dr. Ram concludes: “Using this combination of technology we have shortened our clone selection timelines by two months; in practice we have doubled our clone screening capacity using the same number of scientists, and have observed excellent scale-up predictability. Coupled with the proprietary platform media / feed we have developed for CHO cell lines, we are achieving titer of up to 10 g/L in scale-up. These titers are an improvement on where we were in 2008 when we were achieving 2–4 g/L after lengthy clone selection and optimization and we are sure will contribute to reducing our COGs.”

The Cost of Cleaning

Another strategy for reducing costs that was discussed at the conference was single-use technology both for production and purification of biologics. To show the level of process improvement that can be achieved with single-use technology, Nigel Depledge, Ph.D., staff scientist at Fujifilm Diosynth Biotechnologies, showed an example of a time-in-plant study comparing single-use Sartobind® membrane adsorbers, prepacked chromatography columns and ion exchange columns, which have to be packed, equilibrated, and cleaned. To purify 150–200 L of process material, using the column plus packing and cleaning took 64 hours; the prepacked column took 22 hours and the membrane adsorber three hours. Depledge states: “Single-use technology can decrease plant time for production by up to 95 percent and as a result significantly decreases operating costs.”

To increase biologics production, combining single-use technologies and changing from batch to using continuous processing was mooted as a good strategy of achieving even greater production capacity. According to Konstantin Konstantinov, Ph.D., vice president of late-stage development at Genzyme, Janet Woodcock, the director of the Center for Drug Evaluation and Research (CDER) at the FDA has said the still-existing technology concepts of the 1950s will be abandoned in the next 25 years and bioprocesses will be converted for cleaner, efficient, continuous manufacturing. This encourages companies to consider converting their batch stainless steel plants to more advanced continuous biomanufacturing plants.

For upstream processing, Dr. Konstantinov suggests that perfusion culture can replace batch culture and requires PAT sensors to monitor cell density and automated pumps to be activated to add feed to maintain it at a steady state. He presented data on perfusion culture for biomanufacturing to show that cells need to be maintained at 50–120 million/mL, which operationally means increasing cell density from those traditionally used in batch mode and using a low perfusion rate of 1–2 media volumes per day.

Dr. Konstantinov says: “With continuous perfusion culture you operate at steady state, which reduces product quality heterogeneity as the cells are not under the same stress as they would be when cultured in batch. This means more consistent quality and yield of our monoclonal and nonmonoclonal antibody-based therapies.”

For downstream processing, Konstantinov suggests that continuous processing requires working with multiple smaller columns at a higher binding capacity in lieu of one large industrial-scale column. He presented data to show that using a modified ÄKTA™ system from GE Healthcare operated according to the three-column periodic counter current chromatography (3C PCC) principle linked to a second PCC system involving a membrane adsorber resulted in processing time as fast as 22 hours from media to purified product. According to Dr. Konstantinov this purification normally takes days and the set up is faster because there is not any hold or cleaning steps involved. Using continuous processing instead of batch purification, the use of harvest and clarification tanks is eliminated and the use of buffers and resins are reduced, which in turn reduces costs.

Dr. Konstantinov concludes: “The FDA feedback we have had is that there is no new regulatory constraints to implement this continuous processing concept and currently we are running this at pilot scale in single-use systems. The small volume of the equipment provides the ability to rapidly increase or decrease manufacturing capability as we can number up the upstream and downstream components as and when we need to. However, one issue we have found is, for this process to become truly industrialized, there has to be one software that controls all the systems; currently each part of the process has its own control and this is where a good industrial academic collaboration could help to develop and an overarching control program.”

Future Challenges

To ensure that the links between industry and academia remain strong, the U.K. government is investing in building the National Biologics Manufacturing Centre (NMBC) in Darlington, which alongside the Cell Therapy Catapult will help perform basic research with process development in biologics and cell therapy manufacturing respectively. The Bioprocessing Research Industry Club, an academic-industry research initiative, has also funded 46 academic projects at 32 universities to the tune of over £25 million ($40.8 million) and also funded an additional 28 Ph.D. studentships in collaboration with industry.

Chris Dowle, Ph.D., director of the NBMC states, “We will be up and running by 2015, and the NBMC will have the capacity to support the development of new process technologies and manufacturing routes. We will provide both large and small companies with open access facilities to prove and scale up their process, therefore reducing risk associated with product development. We believe having access to this facility will help position the U.K. as a great place to locate biomanufacturing.”

The U.K. cannot rest on its laurels because there are challenges, and although it produces 5.5 percent of the global biologics portfolio, it remains static in terms of growth. Steve Bagshaw, managing director of Fujifilm Diosynth Biotechnologies UK says, “The U.K. wants to grow its biomanufacturing to a £12 billion (almost $20 billion) industry by 2025, but to do this we need a workforce with the right skill set. Our major problem here is going to be providing enough skilled workers to run our plants, and we need 100,000 more STEM employees every year for the next 10 years just to replace our current workforce and grow marginally across all U.K. industries. So education and even in-house apprenticeships partly funded by the U.K. government may be the answer.” Dr. Bustard adds, “The U.K. was second in the world in terms of biologics pipeline, now it is third behind the U.S. and Switzerland. The U.K. therefore should look to be more ambitious; it has to aim for having a well-trained workforce and implementing the industrialized processes we have seen presented at the Annual bioProcessUK Conference to grow the pipeline considerably if the U.K. wants to remain a prime location for biomanufacturing.”

1 GSK has since confirmed that it is investing £200 million ($326.9 million), some of which will be used to increase biomanufacturing capacity of its antibiotic, Augmentin.

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