Invited Oral Abstract Presentation
On-line control of glucose concentration in high-yielding mammalian cell cultures enabled through oxygen transfer rate measurements
Stephen Goldrick1, Kenneth Lee2, William Holmes3, Marcel Kuiper3, Richard Turner3 and Suzanne Farid4, (1)University College London, London, United Kingdom, (2)MedImmune, Gaithersburg, MD, USA, (3)MedImmune, London WC1E 6BT, United Kingdom, (4)University College London, London WC1E 6BT, United Kingdom
2017 SIMB Annual Meeting and Exhibition
The primary method for glucose control during mammalian fed-batch fermentations, typically involves dynamic bolus glucose additions based on infrequent off-line daily samples. This glucose control strategy results in cells experiencing significant glucose concentration fluctuations which have been previously reported to influence product quality and growth. Current on-line methods to control glucose require expensive process analytical technology (PAT) devices that have proven to be difficult to validate and are not commonly incorporated in industrial biopharmaceutical facilities. This work proposes an on-line method to control and manipulate glucose utilising readily available process measurements. The method has been verified across multiple scales and used varied cell lines. The method generates a correlation between the cumulative oxygen transfer rate and the cumulative glucose consumed. The correlation generates an on-line prediction of glucose that has been successfully incorporated into a control algorithm manipulating the glucose feed-rate. This advanced process control (APC) strategy enables the glucose concentration to be maintained at an adjustable set-point and has been found to significantly reduce the deviation in glucose concentration in comparison to conventional operation. The method has been validated for production of various therapeutic proteins and successfully demonstrated on the micro-(15 mL), laboratory-(7 L) and pilot-(50 L) scale systems.
Invited Oral Abstract Presentation
Antibody fragment production faster and more cost-efficient than CHO: Applying E. coli secretion technology
Dr. Martina Huber, Wacker Biotech GmbH, Jena, Germany and Mark Berge, MedImmune, Gaithersburg, MD, USA
2017 SIMB Annual Meeting and Exhibition
WACKER’s
E. coli based secretion technology (ESETEC
®) was applied to produce the target protein, a recombinant human antibody Fab fragment which was developed by MedImmune to rapidly and specifically reverse the effects of a drug. Administration of relatively high doses of the product made determining a more cost-efficient manufacturing a process objective for this project. While cost-of-goods based on mammalian CHO cells were not optimal, ESETEC
® was the enabling technology for successful production of the Fab. Following a two-month feasibility study, the entire USP and DSP process was developed within nine months and scaled up to WACKER’s 300-L GMP production facility. The highly reproducible manufacturing process delivered 1.15 kg of non-GMP and 2.1 kg of GMP material from a 200-L fermentation scale just 18 months after initiation of the project. In parallel, a second-generation process was implemented with an improved ESETEC
® strain to further increase product titers of the Fab, which was achieved using a definitive screening design approach in USP development.
Invited Oral Abstract Presentation
Break
2017 SIMB Annual Meeting and Exhibition
Invited Oral Abstract Presentation
Advances in adaptive feeding control – a closed loop system to monitor, control, and automate upstream mammalian and microbial feeding protocols
Greg Emmerson1, Sam Watts1, Simon Saxby1 and George Barringer2, (1)Stratophase Ltd, Romsey, United Kingdom, (2)Stratophase Ltd (USA), W Groton, MA, USA
2017 SIMB Annual Meeting and Exhibition
An in-line process monitoring system, the Ranger™, and method for in-situ, real time monitoring and control of nutrient and carbon source feeding in upstream bioreactors and fermentors is described. The Ranger responds to the overall state of the metabolic environment of the process under observation and is highly sensitive to any molecular level perturbation in the process media, such as occurs when a biological process is fed nutrients and carbon sources. An automatic, closed loop adaptive feeding protocol that responds to these real time changes in nutrient concentration in the media then maintains proscribed optimum nutrient concentrations thus permitting faster process development cycles, time to clinic, and the promise of better product quality and titer. The system can actively and independently manage multiple feeds. The system is applicable to all scales of operation from process development to commercial production and is compatible with SUBs. This technology is applicable in microbial, fungal, and mammalian cultures.
Invited Oral Abstract Presentation
Fermentation and strain design for successful scale-up
Michael Japs, Genomatica, Inc., San Diego, CA, USA
2017 SIMB Annual Meeting and Exhibition
Genomatica has established a track record of success in bioprocess development, technology transfer, and scale-up; delivering processes that work commercially the first time. In this presentation we’ll share insights into our validated platform for bioprocess scale-up and bioreactor design. This includes how commercial scale conditions are taken into consideration in the earliest phases of strain design; how to choose fermentation and downstream processing approaches based on metabolic pathway and techno-economic analyses; and designing strains that optimize metabolism and product production under conditions compatible with “at-scale” constraints. The systematic evaluation of process robustness using these unique “scale-down” modeling and experimental methods minimizes scale-up risk and accelerates commercialization timelines for industrial fermentation processes.
Invited Oral Abstract Presentation
Real-time analysis of fermentation and cell culture enabling real-time optimization of profitability
Dr. Joel Sirois, BioIntelligence Technologies inc., Sherbrooke, QC, Canada
2017 SIMB Annual Meeting and Exhibition
Biomanufacturing and bioprocess development suffer from a lack of probes to perform a valuable monitoring of product biosynthesis and its optimization. Many samples have to be taken and analyzed with instruments such as HPLC to have an idea of the kinetics and yields during fermentations and cell cultures. This constrain induces delays and losses. The BioAnalyst is a new instrument collecting data from any source, being probes on a bioreactor, datafile from a HPLC or user entries from manual analysis, and performing real-time calculations to unveil hidden information such as biokinetics. The embedded algorithms push information on personal sets of dashboards for each user enabling them to accelerate decision making and implement real-time optimization. A layer of algorithms is dedicated to translate scientific information to financial information. Users can perform real-time optimization of profitability, identify bad batches and stop them early in the process and project culture behavior over time to anticipate problems and fix them before they happen.
Invited Oral Abstract Presentation
Real-time in-situ fermentation monitoring using generalizable near-infrared assays
Nosa Agbonkonkon, Michael D. Leavell, Daniel Yim, Derek Abbott and Sara Gaucher, Amyris, Emeryville, CA, USA
2017 SIMB Annual Meeting and Exhibition
Accurate measurement of feedstock consumption and product production is required for process development. Traditionally fermentation broth samples are taken and analyzed in batch. This approach is extremely resource intensive and results in a lengthy cycle time. For in-situ product titer monitoring, spectroscopic probes and multivariate data analysis are often used. Current thinking is that large data sets, in the 100’s or 1000’s of data points, are required to make calibration models and that models are not generalizable. This presentation will focus on the development of Near-IR assays for real-time in-situ measurement of product titers utilizing generalizable calibration models, and outline the cost and time savings realized from such an approach.