Kakes of Applikon sees a trend toward more scale-down, with the goal of getting more data more quickly for process development at small scale. “In the old days, you would do screening in shake flasks and try to select the best-producing strain based on a lot of cultures and very little data on the process.”
Now the emphasis is on collecting more data at small scale in shake flasks or microtiter plates to make better data-driven choices. “What we used to do at the 3–15 L scale, we can now do in mini-bioreactors at 50–250 mL scale, and I think in the future will be even smaller.”
Applikon introduced its mini-bioreactor product line in 2011 and, at the time, predicted that “it would cannabilize part of our classic laboratory line that we have been selling for 30–40 years,” says Kakes. “But on the contrary, we are selling a lot of the mini-bioreactors, and it has boosted the sales of the standard lab line. Customers can now go from initial screening done in microtiter plates through the minis and lab scale and up into production scale with either single-use or reusable technology.”
“We see a clear trend for a greater number of bioreactor experiments,” says Zoro of TAP, which offers the ambr™ advanced microscale (10–15 mL) bioreactor system. He identifies cost and other practical issues as limiting factors in the number of parallel benchtop reactors a company can manage. Greater reliance on automated microscale reactors is an increasingly popular strategy for improving bioreactor experiment throughput, according to Zoro.
“Ongoing pressure to decrease time to market and progress more projects within the same teams continues to drive efficiency increases in process development (PD) workflows,” he says.
“Optimized clone screening and process development schemes are increasingly sophisticated, with higher-throughput approaches for early screening and PD tools. These include smaller-scale fed-batch models, greater use of automation, and implementation of microscale reactors, enabling bioreactor design of experiment (DoE) studies for both media and PD.”
“Many biopharmaceutical companies are taking strategic positions in the biosimilar space, building capability in this area either via aggressive (biosimilar) or defensive (biobetter) approaches,” says Zoro. “In either case, key tools required include a scalable early model for clone screening on the basis of product quality and a platform for DoE bioreactor studies to enable rapid optimization of media and process conditions to achieve product quality targets.”
“The past five years in particular have seen increasing demand for parallel bioreactor systems, driven by overall fast growth in the bioprocessing market and ongoing demand for shorter bioprocess development times,” says Kathrin Schmale, Ph.D., strategic marketing manager at DASGIP Information and Process Technology, which manufactures parallel bioreactor systems and was acquired by Eppendorf earlier this year.
“We believe users are looking for all-in-one solutions that combine bioprocessing hardware, process control, and a range of software functionality for data management, analysis, and storage, as well as DoE and automated processing by interconnectivity with third-party lab devices,” says Dr. Schmale.
Despite growing interest in single-use systems for bioprocess development applications, “I feel that there is no shift from glass to single-use bioreactors up to now,” she says.
“This is mainly due to economic factors: in R&D and PD applications, where usually multiple bioreactors have to be operated in parallel, the reduction of setup times correlated with single-use bioreactors does not add up with the current relatively high purchase costs. As long as there is no economic attractive solution available for these applications, the market will choose reusable bioreactors. But I am convinced that we can expect a broader range of single-use solutions with a better price-performance ratio in the near future.”
“Single-use replacements for glass benchtop reactors have had a modest uptake, as the incremental benefits in time saving vs. autoclavable reactors are limited; experimental throughput is not substantially increased. By contrast, use of automated disposable microbioreactors is rapidly becoming standard industry practice, due to improved scalability of results vs. shaken models, combined with improved data consistency and an order of magnitude increase in the number of reactors each scientist can operate, greatly increasing bioreactor experiment throughput,” says Zoro from TAP.
Infors has been expanding its focus beyond the traditional markets of cultivating microorganisms and cells for biopharma applications, adding dedicated technology targeting second- and third-generation biofuels development and photosynthetic applications. Infors developed the laboratory-scale Labfors 5 BioEtOH bioreactor for the second-generation bioethanol sector, allowing for simultaneous saccharification and fermentation.
Mixing of the pretreated lignocellulose slurry prior to enzymatic hydrolysis into a liquid form is a critical and challenging step. “To solve this issue, different kinds of impellers have been developed and a special motor was designed to fulfill both the different needs of the high viscous slurry and the liquid after hydrolysis,” says Daniel Egger, marketing manager, Infors.
To simulate sunlight in the laboratory for photosynthetic applications, the company developed the Labfors 5 Lux illumination system for small-scale algae photo bioreactors. High-powered LEDs produce irradiation as strong as bright sunlight and an irradiation spectrum that approximates sunlight, according to the company. Its Iris parallel bioprocess software can simulate the day/night sunlight curve of the irradiation unit.
Later this year, Infors plans to introduce an automatic cleaning and sterilize-in-place feature (LabCIP) to its Labfors 5 benchtop glass bioreactor line (for sizes 2 L–13 L total volume) and, working with an industrial development partner, has shown that it can be used to double productivity. This will offer a sterilizable option for microbial bioprocessing in a reusable system.
Kakes predicts future growth and standardization in sensor technology. Users still often rely on classic sensor technology because they are not yet confident in the new single-use sensors, in Kakes’ view. To facilitate the transition, Applikon integrates both options into its systems, so users can run a bioreactor initially using classic sensors and then, “when they are confident that the new technology gives the same results, they can switch over to the single-use sensors.”
Looking toward the future, Applikon is developing new types of sensor technology, in particular optical sensors capable of cell-image analysis. During the second half of this year, the company will also introduce a new data-analysis system for easier data integration and processing.
EMD Millipore recently introduced the Mobius CellReady family of single-use bioreactors, which includes 50 L and 200 L sizes. The design of the single-use bioreactor process container includes a rigid base and top panel to simplify and ensure the accuracy of its installation into the stainless steel holder. “We created the base and top panel slots to make the installation process easier,” says Krishnan. Users can only install the bag one way to minimize the chance for errors.
To further enhance the flexibility of single-use systems, EMD Millipore has taken sensors away from the bioreactor and built them into an external loop system that users can configure as needed, reducing the need to customize bags.
Millie Ullah, senior global product manager for bioreactors at Thermo Fisher, highlights two main trends in the bioprocessing market: customer demand for increased standardization and for more components to support process analytical technology (PAT). For PAT the emphasis is on emerging single-use sensor technologies to measure parameters such as pH, dissolved oxygen, metabolites, biomass, and carbon dioxide.
The drive toward standardization is twofold, focusing on connectivity and integration between different products upstream and downstream of the bioreactor, and secondly standardization of the regulatory and quality aspects of supply chain for traceability.
Standardization of system design presents a dilemma. “Although many people want standardization of products, at the end of the day, it does not work for everyone’s process, and in the end we find that customers want to customize to their process specific needs,” says Ullah. Different single-use systems will necessarily have different designs, depending on what type of cell culture processes they are optimized for, such as microcarrier-based cultures or perfusion processes.
While Thermo Fisher has been focusing much of its product development on meeting the varied needs of bioreactor systems, it offers customers the choice of off-the-shelf products with shorter delivery times or of process-specific products tailored to a particular application.
New to the Thermo Fisher bioprocessing line this year is the HyPerforma Single-Use Bioreactor (S.U.B.) Turn-Key (TK) system that combines either a 50 L or 250 L S.U.B. bioreactor with an integrated controller platform composed of PC and Delta V controller systems from Finesse. The company also introduced the second-generation S.U.B., with an updated look and added features.