When expressing protein, there are a variety of expression systems to choose from. CHO is seen as the ‘king’ in the industry, but does it meet all needs of rapid production and budget constraints? This meeting will explore “Optimizing Protein Expression” through understanding and enhancing expression systems, and will especially focus on CHO cells, and other mammalian systems, E.coli and yeasts. Other host systems, such as baculovirus and algae, will be touched on as well.
Along with case studies, the meeting will feature experts who reveal the underlying mechanisms and insights into the varying systems in order to enhance protein expression. Comparing and contrasting systems will also be featured to provide greater understanding of how these systems function within the context of the results achieved.
Final Agenda
Recommended Short Courses*
SC13: Phenotypic Screening Applications and Technologies - Detailed Agenda
SC17: Transient Protein Production in Mammalian Cells - Detailed Agenda
*Separate registration required,
WEDNESDAY, MAY 3
7:30 am Registration and Morning Coffee
8:30 Chairperson’s Remarks
Oliver Spadiut, Ph.D., Principal Investigator, Biochemical Engineering, Vienna University of Technology
8:40 KEYNOTE PRESENTATION: Protein Expression Demands and Demanding Protein Expressions: Protein Sciences the BioPharma Way
David Humphreys, Ph.D., Director and Head, Protein Sciences, UCB NewMedicines
Protein expression in BioPharma can span from small quantity bespoke expression of antigenic proteins for immunisations and assays, large numbers of antibodies to support project discovery needs through proteins for structural studies to therapeutic antibody manufacturing systems. UCB uses a whole range of expression and purification systems from E. coli to mammalian cells to meet project and manufacturing demands. These will be highlighted in a general protein expression context.
9:10 Early Development Strategies for Innovative Therapeutic Molecules Now and Then
Nicola Beaucamp, Ph.D., Head, Process Research, Pharma Research and Early Development, Roche Innovation Center
A number of novel antibody formats have been advanced into clinics by Roche pRED. In order to discover and develop differentiated monoclonal antibodies, Roche’s strategy is based on engineering technologies that bear several challenges for technical development. Examples on innovative therapeutic molecules for multi-pathway-inhibition or specific tumor-targeting will be given.
9:40 Alexion’s Protein Production Process and Platforms: Comparison of Expi293 and ExpiCHO Expression Systems
Tadas Panavas, Ph.D., Associate Director, Discovery Research, Alexion Pharmaceuticals, Inc.
This presentation will showcase how Alexion has implemented streamlined processes across multiple sites to automate our engineering and production of enzyme replacement therapeutics (ERTs) as well as single and multi-domain antibodies. The selection of expression system for antibody therapeutics and ERTs will be discussed, and thorough comparison between Expi293 and ExpiCHO will be presented.
10:10 Coffee Break in the Exhibit Hall with Poster Viewing
10:55 Deletion of a Telomeric Region on Chromosome 8 Correlates with Higher Productivity and Stability of CHO Cell Lines
Corinne Ueberschlag, M.Sc., Fellow, Functional Lead Cell Line Development, Biologics Technical Development and Manufacturing, Novartis AG
A new parental CHO cell line lacking a telomeric region of the chromosome 8 was generated, resulting in significantly improved productivity of the cells after gene transfer as well overall improved production stability of these cells. Combining this cell line with new vector technologies, we have implemented a high performing platform for therapeutic protein production.
11:25 Prediction and Mitigation of Production Instability of Recombinant CHO Cell Lines
Ulrich Göpfert, Ph.D., Principal Scientist, Cell Line & Molecular Development, Roche Innovation Center
During expansion and maintenance, CHO cell lines are prone to production instability, which may be caused by promoter silencing, loss of transgene copies, or post-transcriptional effects. Silencing of recombinant genes may be accompanied by DNA methylation and histone modification. We examined a variety of epigenetic modifications and identified molecular indicators which provide the opportunity to enrich stable producers.
11:55 Lessons Learned Using CHO Transient Systems as an Early Screen for CHO Stable Expression
Barry A. Morse, Ph.D., Principal Research Scientist, Biologics Research, Janssen Research and Development
12:25 pm Applying QbD Principles in Discovery and Early Stage Process Development Facilitates the Scale-Up of Robust Processes
Remi Laliberte, Process Development Manager, Integrated Solutions, Sartorius Stedim North America
There is substantial potential for the research & development space to increase its impact on today’s industry goals through a more integrated approach between discovery and process science.New technologies and services are becoming available that allow for a more seamless integration of discovery work, CQA’s, design spaces, process development, process/product quality control. The integration of those technologies across the biopharma drug development cycle will allow for substantial efficiency gains/time savings getting molecules to clinic.
12:55Luncheon Presentation: New Tools for Screening & Harvesting Solutions for CHO & HEK293 Cells, for Both Transient and Stable Cell
Sam Ellis, Vice President & Biochemist, Thomson Instrument Company
Evaluation of different transfection tools, product quality, and titer for both CHO and HEK293 cell lines. Data will be presented on techniques and technology that mimic large-scale bioreactors in non-controlled devices from 1mL-3L. Technologies presented include well plates and culture tube systems with incorporated filtration methodology. A new direct harvesting technique will also be introduced that eliminates centrifugation while maintaining 0.2um sterile filtration. All of these tools will be presented with case studies from scientists.
1:25 Luncheon Presentation: Difficult to Express Proteins: Novel Plasmid Technology to Significantly Increase Product Yield in CHO Cells
Marco Cacciuttolo, Ph.D., Director Operations, Batavia Biosciences
Yield is still an area that requires significant improvement for many promising recombinant protein and antibodies. Especially in the field of biosimilars and orphan diseases, the ability to manufacture products cost-effectively could have a profound impact on the interest of the industry to develop safe, affordable and efficacious products. Novel plasmid technology enables rapid generation of stable, CHO cell lines able to provide at least 10-fold more product per cell.
1:55 Session Break
2:10 Chairperson’s Remarks
Tadas Panavas, Ph.D., Associate Director, Discovery Research, Alexion Pharmaceuticals, Inc.
2:15 Codon Influence on Protein Expression in E. coli Correlates with mRNA Levels
John Hunt, Ph.D., Professor, Biological Sciences, Columbia University
We have developed new computational gene-design algorithms based on multiparameter modeling of results from 6,348 experiments employing bacteriophage T7 polymerase to drive protein overexpression E. coli. Our analyses define the relative influence of mRNA sequence parameters including predicted folding energy, base composition, and codon usage. Our new codon-influence metric differs significantly from prior expectations and permits generation of diverse synonymous gene sequences that consistently drive high level protein overexpression.
2:45 How Induction Impacts Inclusion Body Properties and Inclusion Body Processing in E. coli
Oliver Spadiut, Ph.D., Principal Investigator, Biochemical Engineering, Vienna University of Technology
I will show our ongoing work, where we 1) understand the effect of lactose/IPTG induction on E. coli physiology and productivity, 2) demonstrate the effects of induction on inclusion body properties (size, amount, density, purity), and 3) finally show how induction and consequent inclusion body properties affect the subsequent inclusion body processing (wash, solubilization, refolding).
3:15 Expression Flexibility at the 2-12L scale: SB10 Orbital-Shaken SUB for High Density Cultivation in Multiple Cell Types
David Laidlaw, CEO, Kuhner Shaker, Inc.
Batch, fed-batch and perfusion protein production may be accomplished using a number of platforms. Production simplicity is achieved when small scale results predict those of the bench scale bioreactor. Here we present data from users of the orbital-shaken SB10 bioreactor with CHO, Sf9, AGE1.CR and moss cells (P. Patens).
3:30 The 3rd Generation Strep-tag® System – Superior Performance in Protein Purification and Assays with Strep-Tactin®XT
Uwe Carl, Ph.D., Head, Protein Production, Strep-Tag Products and Proteins, IBA Lifesciences
IBA is focused on building a comprehensive product portfolio around its proprietary Strep-tag® technology to provide solutions for protein production and assays, including cloning, expression, purification and immobilization. Especially our 3rd generation Strep-tag® system is superior to other systems due to its extreme high affinity and still reversible binding.
3:45 Refreshment Break in the Exhibit Hall with Poster Viewing
4:45 Problem-Solving Breakout Discussions
These interactive discussion groups are open to all attendees, speakers, sponsors, & exhibitors. Participants choose a specific breakout discussion group to join. Each group has a moderator to ensure focused discussions around key issues within the topic. This format allows participants to meet potential collaborators, share examples from their work, vet ideas with peers, and be part of a group problem-solving endeavor. The discussions provide an informal exchange of ideas and are not meant to be a corporate or specific product discussion. Pre-registration to sign up for one of the topics will occur a week or two prior to the Event via the App.
Common Issues with Transient Protein Production
Richard Altman, MS, Scientist, Protein Technologies, Amgen
Henry C. Chiou, Ph.D., Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific
Dominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc.
Scalable and rapid transient protein production in mammalian cells continues its evolution as an integral part of the biotherapeutic drug discovery process. We discuss the common issues facing researchers as they try to meet an expanding demand for transiently-produced recombinant protein.
- What are the current challenges to transient protein production?
- What are the keys to optimizing expression?
- How do we optimize the whole protein expression process?
- What scale of expression and level of throughput are commonly being used?
- What cell line should we use and when?
- Characterization of transiently-produced proteins
Transient CHO/HEK vs. Stable CHO?
Sam Ellis, Vice President, Thomson Instrument
- Can you get high-quality titers in 2 weeks or less?
- Transfection reagents, vessels, and media
- how to accomplish high titers in less time, and meet your group’s needs
- Group discussion on how to accomplish this for reagent and therapeutic uses.
Parallelized Protein Purification
Allan Matte, Ph.D., Senior Research Officer, Human Health Therapeutics, National Research Council Canada
- Equipment and technologies to achieve parallelized protein purification at various scales
- Higher throughput, small-scale protein purification using liquid handlers for example
- Integration of auto-sampler capabilities with purification platforms to increase throughput
5:45 Networking Reception in the Exhibit Hall with Poster Viewing
7:00 End of Day
THURSDAY, MAY 4
8:00 am Morning Coffee
8:30 Chairperson’s Remarks
Nicola Beaucamp, Ph.D., Head, Process Research, Pharma Research and Early Development, Roche Innovation Center
8:35 Pathway Engineering for Customizing Eukaryotic Protein Glycosylation
Nico Callewaert, Ph.D., Director, VIB Medical Biotechnology Center, Ghent University
Glycosylation is the most common post-translational modification on biopharmaceuticals produced in eukaryotic hosts. Developments over the past decade have enabled to achieve higher levels of control over the glycan structures on biopharmaceuticals, allowing to customize these structures for particular therapeutic functionality. In this talk, I will discuss Pichia pastoris GlycoSwitch (TM) and mammalian cell/yeast/plant GlycoDelete technologies, as well as position these novel systems within the available range of biopharmaceutical expression hosts.
9:05 Implementing High Temperature Short Time (HTST) Viral Control Strategy in Large-Scale Biotherapeutics Manufacturing
Min Zhang, Ph.D., Director, Manufacturing Sciences and Technology, AstraZeneca/MedImmune
Adventitious viral contamination poses a potentially devastating risk to Biotherapeutics Manufacturing. The chance may be low, yet holds the potential to create a huge impact including the financial cost to business that could ultimately affect patients. As a single, critical component of the overall viral control risk mitigation strategy, HTST has been accepted industry-wide as an effective preventative step for viral risk mitigation. This presentation will highlight an effective HTST implementation strategy at the AstraZeneca Frederick Manufacturing Center (FMC) that has been established with collaborations with cross-functional organizations in AstraZeneca.
9:35 Accelerating Development Timelines Using Scalable Delivery Platform for Transient and Stable Protein Expression
James Brady, Ph.D., Vice President, Technical Applications and Customer Support, MaxCyte, Inc.
Transitioning early stage discovery with later stage development is critical for efficiently moving therapies into the clinic.MaxCyte’s electroporation-based delivery platform enables high titer expression of antibodies and other proteins in cells that are relevant to biomanufacturing. We present data using MaxCyte electroporation for gram scale transient antibody production followed by rapid and streamlined migration to stable cell lines. Finally, we discuss means for targeted genomemodification for generation of cells engineered for maximum productivity.
10:05 Coffee Break in the Exhibit Hall with Poster Viewing
11:05 Application of a Baculovirus Platform for the Production of Vaccines
Indresh K. Srivastava, Ph.D., Vice President, Product Realization, Senior Project Manager, Protein Sciences Corp.
In this talk, I will present the data on the production, characterization and immunogenicity of our FDA-approved Trivalent and Quadrivalent Influenza Vaccines, and strategy to perform process improvements for yield improvement. Will also present the data on ZIKV vaccine development program.
11:35 Production of Recombinant Proteins in Eukaryotic Green Algae: Growth of a New Green Sector
Miller Tran, Ph.D., Associate Director, Triton Algae Innovations, Inc.
At Triton Algae Innovations, we focus specifically on the chloroplast of eukaryotic green algae. Chloroplasts house complex molecular chaperones and protein disulfide isomerases that allow them to efficiently and economically produce a large class of proteins. Another interesting facet of algae is their ability to be consumed. This trait effectively eliminates the costly process of protein purification. However, moving algae from a system used for biofuels to one used to produce recombinant protein required a shift in production strategies and a development of a novel suite of expression tools to facilitate a new growth and production strategies. At Triton, Dr. Tran has pioneered the development of these expression vectors and the production of a large class of proteins for use in the gut health industry.
12:05 pm Pichia-Based Recombinant Proteins: From Difficulty to Development
Chandrasekhar Gurramkonda, Ph.D., Research Assistant Professor, Chemical, Biochemical and Environmental Engineering, University of Maryland
Pichia pastoris emerged as a next door expression organism for many scientists and engineers. Because of its ability to reach high-cell densities, it can grow in simple buffer and GRAS host for the expression of recombinant proteins such as disulphide bonded proteins, oligomeric forms and proteins with its cognate partners in very high quantities by multicopy integration.
12:35 End of Optimizing Protein Expression