CCDWC 2017 Day 2
Upstream – CRISPR / Cell line development
Upstream - Media and process development
Downstream – Advances in chromatography and novel purification methods
Formulation
09:00
From thousands of clones to the final one
- CHO cells are the most widely used host for large-scale production of recombinant therapeutic proteins
- In order to identify the most suitable lead cell lines Novartis started to introduce a development process that comprises thorough early candidate profiling focusing on biophysical properties of candidates, followed by a cell line development process
- applying an efficient selection strategy in combination with process-tailored automation for clone imaging, clone handling and selection
09:00
Cell line development and process development for biosimilars
- Definition of Development goals
- Choice of cell lines
- Quality Attributes during Process Development
09:00
Continuous chromatography – challenging bioprocess development problems
- Continuous operation of polishing chromatography
- Robustness and fault diagnosis of continuous chromatography
- Integration of continuous steps
09:00
Lyophilization in Dual Chamber Cartridges – from Energy Transfer to Process Control
- Process challenges during freeze-drying in DCCs
- Selection of adequate holder devices
- Process design and control with the “DCC-LyoMate procedure”
09:20
CHO cell factory engineering, for working towards improved production of therapeutic proteins
- CHO cell line engineering and therapeutic protein production
- CRISPR/Cas9-mediated genome engineering technologies
- Targeted integration to facilitate controlled and reproducible cell line engineering
09:20
Increase yields via media optimisation toolbox and process parameters improvement
- A custom media optimization approach was designed for use with the ambr® 15 microbioreactor system
- Fed-batch processes were developed for multiple case studies for cell lines producing a variety of antibodies
- Media optimizations and improvements in peak viable cell density and titer will be compared between culture systems and case studies
09:20
Challenging biomolecule purifications resolved with innovative selectivities
- Novel antibody formats like fragments or non-IgG molecules, lack a strong protein A binding site facilitating efficient capture and purification in a protein A affinity based platform.
- New developments in ligand technology provi10:00de chromatographic materials with unique selectivity as well as high capacities (ligand grafting)
- Discussion will demonstrate the development of a purification platform for non-IgG type antigen binding biopharmaceuticals using these innovative selectivities
09:20
Mabs aggregation due to mechanical stress during fill & finish process
- Mixing: classical bottom agitation may induce very high stress for particular fragile Mabs, and this is the reason why technology of agitation by levitation is spreading in Biotech processes . Very simple lab-scale studies can demonstrate how even low contact between a fix surface and a mobile agitator generate high amount of particulates.
- Filling: in a same manner, piston pump filling implies contact and movement between two stainless steel parts and generates significant quantity of particulates, comparatively to less stressful technologies like peristaltic pump or time-pressure.
- The potential root cause of these Mab degradations occurring during mixing with contact and piston pump filling will be discussed: shear stress, abrasion and formation of material particulates, etc
09:40
Rapid cell line development and efficient genome modification using scalable electroporation
- Flow electroporation transfection technology enables highly efficient loading of plasmids, messenger RNA and other molecules into cell lines and primary cells at scales ranging from <1e6 to 2e11 cells
- Discuss how the high efficiency and viability allows generation of stable pools and clones with reduced time and effort
- Share data on efficient targeted modification of mammalian genomes for cell therapy and protein expression using CRISPR/Cas-9 and zinc finger nucleases
Characterisation of the phosphoproteome of recombinant Chinese hamster ovary (CHO) cells to improve bioprocess-relevant phenotypes
- Phosphorylation is a key post-translational modification, playing a crucial role in regulating many cellular processes including transcription, translation, signaling, etc
- The impact of the phosphoproteome on bioprocess performance is poorly characterized in recombinant CHO cells
- We have identified over 700 differentially expressed phosphopeptides using quantitative label-free LC-MS/MS phosphoproteomic analysis in conjunction with IMAC and TiO2 phosphopeptide enrichment strategies, following a reduction in culture temperature (temperature shift)
- Phosphoproteins have the potential to be cell engineering targets to improve bioprocess-relevant phenotypes
09:40
3D printing of chromatographic columns
- 3D printing as a tool to revolutionize the design chromatographic columns
- Closing the loop between experiments and models
- Direct capture of proteins contained in solid-laden streams
09:40
Lyophiliser characterisation: determination of equipment heat and mass transfer properties to support scale-up and technical transfer of lyophilised biopharmaceuticals
- Lab and commercial scale determination of vial heat transfer coefficient and sublimation behaviour
- Predicting primary drying conditions in different lyophilisers
- Design space generation
- Utilising knowledge of equipment and product to support manufacturing
10:00 Networking Break
10:45
Toolbox for Cell Line Development – Next-Generation Cell Line Development Technologies
- CHO cells are the most widely used host for large-scale production of recombinant therapeutic proteins.
- A novel toolbox of vector elements, selection marker and novel engineered CHO cell lines were developed which results in combination in significant increase of titer and improved product quality.
- We have identified a key protein severely affecting the quality of non-antibody format therapeutic proteins.
- Elimination of this protein via novel targeted gene disruption tools resulted in significant increased improved product quality.
10:45
Recent innovations in liquid media preparation
- Making liquid large volume media preparation more efficient
- Recent innovations in liquid media preparation
- De-bottlenecking large volume liquid media preparation
10:45
Excipient considerations in lyophilised protein formulations
- Commercial protein formulations: a review of excipients
- Impact of arginine on the freeze drying performance of protein formulations
Downstream - PAT, process modelling, monitoring and control
11:05
Beyond GS: Using different genome editing techniques to generate improved commercially available production cell lines
- Genome engineering has significant potential to improve our ability to manufacture biotherapeutic products
- CRISPR technology can be applied to perform rapid proof of concept edits
- After validation, Horizon uses rAAV technology to generate improved CHO cells for commercial release with a clear IP position
- Here we present data on edits that build on our GS null cells to create our next generation cell line platform
11:05
Proteomic identification of bioprocess markers for cell stress and cell death during perfusion culture
- Lab-scale model system for apoptotic and necrotic CHO
- Screening and targeted LC-MS workflow for the identification of protein viability markers from apoptotic and necrotic CHO
- Method application to samples from biomanufacturing
11:05
Applications of Process Analytics in Continuous Downstream Processing
- Conti flow manufacturing can have an enormous impact on manufacturing in terms of quality, quality and cost
- Moreover it is possible to design conti flow systems that are inherently more controllable than batch processes
- To take advantage of these control possibilities requires us to develop robust and reliable real-time process measurements
End of Formulation, Fill & Finish Track
11:25
miRNA engineering of CHO host cell lines facilitates production of difficult-to-express proteins to increase success in cell line development
- MicroRNAs (miRNAs) have emerged as potent cell engineering tools to improve bioprocess performance of CHO cells. We established a novel engineered CHO host cell line constitutively expressing a pro-productive miRNA in order to enhance overall bioprocess performance.
- Novel miRNA engineered host cells were tested in two independent CLD campaigns using two different mAb candidates including a difficult-to-express antibody. Production clones derived from target miRNA overexpressing CHO host cells outperformed non-engineered control cells and exhibited substantially increased product yields without compromising product quality.
- Our results demonstrate that host cell engineering using miRNAs represents a promising tool to overcome limitations in industrial bioprocess development especially regarding challenging proteins.
11:25
Discussion: Media and process discussion and extended Q&A
- Discussion to talk about media and process development across the cell culture industry
- Key challenges that companies and academics are facing
- New movements in this area
- Extended Q&A to the speakers
11:25
A novel toolbox for monitoring inclusion body processing
- Novel toolbox comprising HPLC and chemometric tools to monitor the complex steps in a typical inclusion body process
- Toolbox allows the analysis of these steps within minutes after sampling, thus allowing decisions when a unit operation is done
- Reduces overall process time and costs, and also minimises the risk of product loss
Upstream - Innovation showcase
11:45
Benefits of a platform technology approach for the generation of production cell lines and processes
- Description of CHO based platform technology including essential key components
- Description of Cell line development process and selection strategy to select for high
11:45
The C1 expression system – A journey to displace CHO cells with breakthrough cell expression
- Producer of enzymes and proteins using a proprietary and patented expression system based on the Myceliopthora thermophila fungus, nicknamed C1.
- A hyper-productive fungal expression system used to develop & manufacture large quantities of desired proteins at industrial scale at significantly lower CapEx and OpEx costs.
- Can produce Vaccines at high level and with excellent immune response - Dyadic has also displayed the ability to easily express mAb’s.
11:55
Getting the full picture: Addressing Protein Stability by nanoDSF
- How to make better informed decisions earlier in the biologics development workflow
- Explore the modern alternative to DSC: 100x faster, 40x less sample
- Predicting stabilities: thermal and chemical unfolding, melting points and deltaG, aggregation behaviour
12:05 Networking Lunch
13:00
Ultra-deep next generation mitochondrial genome sequencing reveals widespread heteroplasmy in Chinese hamster ovary cells
- First mitochondrial genome sequence of the Chinese hamster
- Widespread heteroplasmy across a panel of Chinese hamster ovary cell lines
- At least one heteroplasmic variation/cell line which resulted in an amino acid change
- Potential impact on CHO cell metabolism
13:00
Use of PAT for process development and control – Toward process digitalization
- Overview on statistical tools for process screening, development, monitoring and control. Where can we save money and how to use such tools for quality management?
- Use of online monitoring and supervisory control for process development. Is it too early?
- Introduction to advanced methods for process development: hybrid models. How to effectively manage knowledge during process development?
- Future perspectives on process digitalization
Michael Sokolov, PhD Candidate, ETH Zurich
13:20
microRNAs for optimized production: towards a deeper understanding of underlying mechanisms
- miRNAs represent promising tools for cell engineering of biopharmaceutical production cells
- Easy-to-use methodology makes miRNAs feasible for usage in production process
- Detailed analyses of underlying mechanisms of action within cells can reveal pathways mediating improved phenotypes
13:20
A holistic approach to analytical support, for the biotech industry
- Growing demand for new analytical tools;
- Increasing infrastructure costs;
- Subcontracting of analytical work
Upstream – Continuous manufacture
13:40
Update on ballroom concept facilities for continuous manufacture
- Status and limitations of continuous processing
- Status implementation of disposables and closed processing concept
- Ballroom plant design for continuous processing
14:00
USP/DSP integration
- Upstream strategies that shift the focus from high titer only to titer in respect to the processability in the downstream
- Increasing titer lead to even more increasing side components, making the downstream very complex and therefor the whole process more expensive than a medium titer with good to separate side components
- Biothermodynamics and online control strategies for the fermenter
14:20 Networking Break
Closing plenary: Integrated processing
15:00
Development and characterization of integrated processes using data science tools
- Identification of interaction effects between unit operations
- Similarity analysis of integrated processes for technology transfer
- Root cause analysis for bioprocesses
- Solutions for process characterization and process validation along the new validation guideline Annex 1 and CPV
15:20
A financial perspective to integrated bioprocessing – Can we make this cost effective?
- Overview on integrated continuous bioprocessing, from an operational and financial perspective
- Cost of good analysis and comparison to standard batch processes. Where do we stay and where should we focus
- Investment (net present value) analysis on integrated process for multi-drug production plants: do we really lose in flexibility?
- Future perspectives: How can industry work together a holistically integrated process? What is still needed to exploit this technology?
15:40
Panel: Integrated processes - how can we achieve this?
- Overview on integrated continuous bioprocessing
- What people are doing to incorporate integrated bioprocessing
- Why integrated processing is important
- What we can look for in the future
16:20
Chair’s closing remarks and thankyou from Terrapinn
16:30 End of Conference
last published: 17/Feb/17 11:15 GMT
Organiser
