Cambridge Healthtech Institute ’s 2nd Annual
Optimising Cell Culture Technology
Enhancing Knowledge for Growing Cells
19-20 March 2019
The ubiquitous task of culturing cells is recognised as an established technology; yet, meeting the demands of industry pushes the need to increase titre, and decrease the time and resources necessary to meet project goals. The Optimising Cell Culture
Technology conference examines strategies to streamline production while ensuring bottom-line quality. The conference will include a session focused on culturing CHO cells (Chinese Hamster Ovary cells), as well as discussions of breakthrough technologies
and analytical approaches that support the tasks at hand and lead to greater productivity. CRISPR, modeling and computational insights will be discussed in the continuing efforts to improve yield.
Short Courses |
Training Seminars |
Tuesday, 19 March
7:00 Registration and Morning Coffee
8:25 Chairperson’s Opening Remarks
Kerstin Otte, PhD, Professor, Institute of Applied Biotechnology (IAB), Pharmaceutical Biotechnology
(PBT), Biberach University of Applied Sciences
8:30 Should the Dynamic Nature of Process Operation Be Reflected by the Design Space Representation/Characterization?
Moritz von Stosch, PhD, Senior Manager, Drug Substance, Technical R&D, GSK Vaccines
The majority of design spaces described in publications follows a “static” statistical experimentation and modeling approach. Given that temporal deviations in the process parameters are of a dynamic nature, static approaches do not suffice.
I will discuss alternative forms of design space representations and illustrate the limitations of the predominantly applied static approach. These approaches may create an opportunity to integrate process characterization, process monitoring and
process control strategy development as defined in the Quality by Design workflow.
9:00 Towards Model Predictive Control of Cell Culture Bioprocesses
Gerald Striedner, PhD, Associate Professor, Biotechnology, University of Natural
Resources and Life Sciences, Vienna (BOKU)
Today, quality by testing is still the gold standard in bioprocesses. The production process is fixed and tightly specified to guarantee a constant product quality. However, out-of-specification events often result in batch rejections. Therefore,
we applied intensified Design of Experiment (iDoE) to quickly screen a defined design space of a CHO fed-batch process. Hereby, a hybrid model is created that is enhanced by the data. This model will be used in the future for model predictive
control to avoid batch rejections.
9:30 Digital Twins for Process Robustness and Control Strategies
Christoph Herwig, PhD, Professor and Head, Biochemical Engineering, Chemical Environmental
and Bioscience Engineering, Technical University of Vienna (TU Wien)
Digital twins get increased attention due to novel enablers of digitalization, but also due to regulatory encouragements such as ICH Q12. But how to use them for fulfilling industrial needs, such as achieving higher process robustness and smooth accelerated
process validation? The contribution introduces concepts and case studies in which digital twins are deployed for smart experimental design and model-based control.
10:00 Grand Opening Coffee Break in the Exhibit Hall with Poster Viewing
10:45 Intensified Design of Experiments – An Approach to Reduce Bioprocess Development and Characterization Times
Mark Dürkop, PhD, Project Leader, Biotechnology, University
of Natural Resources and Life Sciences, Vienna (BOKU)
One of the bottlenecks in the development of innovative biopharmaceuticals is found in endless process development and characterization times. Within this work, we compared classical Design of Experiments (DoE) with an intensified DoE approach
applied on an E. coli fed-batch process. While the screening of a classical DoE required 29 weeks, the intensified characterization was finished within 10 weeks unveiling both the best process conditions and dynamics. The generated
model can further be applied for process control.
11:15 Media Development for Intensification of Seed Train Expansion Including N-1 Perfusion
Luis Fernando Ayala Solares, MSc, Scientist, Perfusion Systems, Cell Culture Media Development, Upstream R&D, Merck KGaA
Seed train expansion as well as the media composition used during the cell culture steps play a critical role for biopharmaceutical production. We could show that the right combination of media, specially designed for their purposes, in a seed train including a perfused N-1 step can increase productivity in the final production step. This indicates that the specific companion media combinations can increase productivity gains with these intensified process formats.
11:45 Next Generation Single-Cell Dispensing in Cell Line Development
Thomas Kretschmann, Field Application Specialist, cytena
Cytena’s single-cell deposition technology isolates single, living cells automatically in 96- and 384-well plates in a documented, viable and pure workflow. The single-cell printer and the new x.sight instruments provide assurance of clonality,
high cell viability, no cross-contamination by the use of disposables and are easy to use.
12:00 Continuous In-Line Cell Counting and Infection Kinetics Monitoring, a Case Study
Sean Case, Scientist, Process Cell Culture: Upstream Scale Up Group, Novavax
Inline, label-free and in real time viral infection kinetics with fed batch process including two different recombinant protein targets; comparability at 10 L vs 50 or 200 L scales to demonstrate viral infection kinetics similarity for small
scale model qualification (SSMQ) and the impact on downstream processing.
12:15 Luncheon Presentation: Generating Production Cell Lines with Superior Titers and 99% Monoclonality in Under 1 Week
Anupam Singhal, PhD, Senior Manager, Technology Development, Berkeley Lights
12:45 Session Break
13:30 Chairperson’s Remarks
Michael Butler, PhD, CSO, Research, National Institute of Bioprocessing Research
& Training (NIBRT)
13:35 KEYNOTE PRESENTATION: Proteomic and Phosphoproteomic Characterisation of Growth of Recombinant Chinese Hamster Ovary Cells
Paula Meleady, PhD, Associate Director, National Institute for Cellular Biotechnology, Dublin City University
Phosphorylation is a hugely important post-translational modification, playing a crucial role in regulating many cellular processes. We describe the proteomic and phosphoproteomic characterization of CHO cells during growth in culture and
in cells subject to temperature shift. Using advanced LC-MS/MS techniques, we have characterised >8000 CHO-specific phosphorylation sites and identified >1500 differentially expressed phosphopeptides in these studies. Phosphoproteins
have the potential to be cell engineering targets to improve efficiency of recombinant protein production.
14:05 Engineering CHO Cell Lines for the Production of Hard-to-Produce Proteins
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk
Foundation Center for Biosustainability, Technical University of Denmark (TU Denmark)
Using our high-throughput cell line engineering platform, we have engineered CHO cells that are able to produce therapeutic proteins that previously were not possible to produce in CHO cells. This approach may result in improved therapeutic
proteins with better biological properties such as increased half-life, improved activity, etc.
14:35 Difficult to Express Proteins: Identification of Intracellular Production Bottlenecks in CHO Cells
Kerstin Otte, PhD, Professor, Institute of Applied Biotechnology (IAB), Pharmaceutical
Biotechnology (PBT), Biberach University of Applied Sciences
With the advance of complex biological format proteins, mammalian expression systems often show low performance. Determining factors may be haltering of heterologous proteins within the different cellular compartments disturbing transport
or secretion. Here we present a streamlined microscopy-based methodology for CHO production cells investigating rate-limiting steps in production organelles, which is also applicable for automated applications in industrial cell line development
processes. Characterisation of identified haltering cellular structures will enable engineering approaches for optimized cellular production.
15:05 Refreshment Break in the Exhibit Hall with Poster Viewing
15:45 A Genetic Engineering Strategy for the Improvement of Chinese Hamster Ovary (CHO) Cell Line Development and Performance
Paul Kelly, PhD, Postdoctoral Research Scientist, Cell Engineering Lab, National Institute
for Bioprocessing Research and Training (NIBRT)
Chinese hamster ovary (CHO) cells remain the dominant production host for recombinant therapeutic proteins within the biopharmaceutical industry. Further enhancing their ability to manufacture these complex therapies in large quantities as
well as shortening cell line development timelines has been the focus of CHO cell biologists over the past 3 decades. Genetic engineering strategies using microRNAs (miRNAs) to augment CHO cell bioprocessing performance have proved promising
but require a greater deal of understanding.
16:15 In silico Culture Media Design Based on Prior Metabolic Knowledge
Rui Oliveira, PhD, Associate Professor, Systems Biology and Engineering (SBE), Chemical
Engineering & Biochemistry, Universidade Nova de Lisboa
In this talk, a rational methodology to design culture media composition based on biochemical networks is presented. The method consists of a systems approach to generate hypothesis on the concentrations of organic compounds, mineral macro-
and microelements in culture media using a priori knowledge on the metabolic network of the target cells. Although not completely eliminating the need for experimental testing, this rational approach
significantly reduces the experimental burden in comparison to statistical DoE-based methodologies.
16:45 Breakout Discussion Groups
This session provides the opportunity to discuss a focused topic with peers from around the world in an open, collegial setting. Select from the list of topics available and join the moderated discussion to share ideas, gain insights, establish
collaborations or commiserate about persistent challenges. Then continue the discussion as you head into the lively exhibit hall for information about the latest technologies.
Process Robustness, Challenges and Validation Strategies
Moderator: Christoph Herwig, PhD, Professor and Head, Biochemical Engineering, Chemical Environmental and Bioscience Engineering, Technical University of Vienna (TU Wien)
- How to judge that a process is robust?
- Is perfusion a robust process
- How to demonstrate manufacturing capability inside of CPV by data science tools
- How to find root causes for process variability
Problem Solving through Genome Engineering
Moderator: Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark (TU Denmark)
- Which problems can and which cannot be solved by genome engineering?
- Which genome engineering tools are being used?
- Which problems have been solved?
Bioprocessing Data Science –Where are the Training Gaps and What will be the Perceived Benefits?
Moderator: Ronan O’Kennedy, PhD, Consulting Bioprocess Specialist, ROK Bioconsulting
- What tools do we need to efficiently leverage and maximise bioprocess data value?
- What are the perceived benefits and possible business case?
- What are the priority training gaps that need to be addressed?
17:30 Welcome Reception in the Exhibit Hall with Poster Viewing
Training Seminars |
Wednesday, 20 March
8:00 Registration and Morning Coffee
8:25 Chairperson’s Remarks
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark (TU Denmark)
8:30 Monitoring the Metabolic State and Productive Capacity of Cells during a Bioprocess
Michael Butler, PhD, CSO, Research, National Institute of Bioprocessing Research
& Training (NIBRT)
Maintenance of cell viability during a bioprocess is crucial for productivity and product quality. The traditional approach involves off-line testing using dye exclusion at fixed points throughout the process. Novel methods of on-line digital
holography, dielectric spectroscopy and impedance flow cytometry can provide a more complete picture of the state of cells during a bioprocess. Such methods compare well with off-line dye exclusion methods and conventional flow cytometry.
9:00 Triple Benefit of BHRF1-Modified Hybridoma Cells: Robustness, Faster Growth And More Balanced Metabolism
Martí Lecina Veciana, PhD, Associate Professor, Bioengineering, Institut
Químic de Sarrià, Universitat Ramon Llull (URL)
BHRF1, a protein with homology with the anti-apoptotic protein Bcl-2, was transfected in hybridoma cells. Besides blocking the cytochrome C secretion, and thus more robust cells against apoptosis were obtained, BHRF1 somehow promoted the influx of pyruvate into the mitochondria. Thus, lactate generation/secretion was reduced (1084 vs 2355 [nmol/mg·h-1]) and higher amounts of ATP produced, resulting in higher cell growth rates (0.040 vs 0.027 h-1) and more efficient glucose and main amino acids consumption. Overall, final cell density was doubled using identical media.
9:30 Process Intensification
David Brühlmann, PhD, Manager, Biotech Process Sciences Technology
& Innovation, Merck Healthcare
Process intensification in upstream bioprocesses to enhance cell culture productivity has become more frequent. This presentation provides insight into the development of an intensified fed-batch platform process, which enables increased
process efficiency with a smaller footprint. Examples include the production of various monoclonal antibodies and a fusion antibody, showing 2-4x titer increases with comparable product quality to classical fed-batch processes.
10:00 POSTER SPOTLIGHT
Involvement of Glutathione during the Production of Recombinant Protein in CHO Cells
Valentine Chevallier, PhD, Associate Scientist, Cell Culture Process Development, UCB
10:30 Coffee Break in the Exhibit Hall with Poster Viewing
11:15 Chairperson’s Remarks
Manuel Carrondo, PhD, Professor of Chemical and Biochemical Engineering, FCT-UNL; Vice President, IBET
11:20 Bioprocessing Innovations in the Era of Clinical Acceleration and Process Intensification
Stefanos Grammatikos, PhD, Vice President, Head, Biotech Sciences, UCB Pharma
Current trends in clinical development acceleration and bioprocess intensification impose an unprecedented compression of CMC development timelines and new bioprocessing challenges downstream of the cell culture bioreactor. In this
talk I will present a series of innovations we have introduced, some incremental and some potentially disruptive, in an effort to avoid further complications while rising to the latest challenges of bio CMC development and bioprocessing.
11:50 Opportunities and Challenges in CAR T Manufacturing
Markwin Velders, PhD, Vice President, Operations, Managing Director, Kite Pharma EU B.V.
Update on the status of CAR-T development for use in the treatment of cancer. The success story of this paradigm shift and the challenges and opportunities that lay ahead for this therapy will be presented and discussed.
12:20 Close of Conference
12:30 Bridging Luncheon Presentation: Next Generation Technologies that Yield Higher Quality CHO Clones and Intensified Gene to GMP Timelines
Alison Young, PhD, Staff Scientist, Mammalian Cell Culture, FUJIFILM Diosynth Biotechnologies
Should biomanufacturing companies rely on well-known, traditional solutions to cell line development with extended timelines, or invest in new technologies with the potential for greater productivity and speed? In this study we demonstrate how to leverage (i) host cell line directed evolution strategies to improve bioprocess relevant phenotypes and increase mAb titres up to 2-fold; and (ii) key technology enablers that allow intensification of cell line development timelines from 25 weeks to 10 weeks.