2016 Archived Content
Cambridge Healthtech Institute’s Ninth Annual
Optimising Expression Platforms
Meeting the Demand for Recombinant Antibodies
2-3 November 2016 | EPIC SANA Lisboa Hotel | Lisboa PORTUGAL
The utilisation of recombinant antibodies for basic research, clinical diagnostics and therapy continues to expand. Consequently, the efficient expression and production of these valuable biomolecules face challenges in improving their quantity and quality
while minimising time and cost. To meet these demands, an increasing variety of recombinant production platforms are being developed. Unfortunately, there is no “universal” production system which can guarantee high yields of recombinant
antibody, particularly as every antibody-based molecule itself causes its own issues in terms of expression.
The Optimising Expression Platforms conference offers comparisons, evaluations and solutions that enable researchers to efficiently express the recombinant antibody of their choice.
Final Agenda
Day 1 | Day 2 | Speaker Biographies | Download Brochure
WEDNESDAY 2 NOVEMBER
07:45 Registration and Morning Coffee
08:30 Chairperson’s Remarks
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, Novo Nordisk Foundation Center for Biosustainability (CFB), DTU Biosustain, Technical University of Denmark
08:35 KEYNOTE PRESENTATION:
Animal Cells in Bioreactors for Production of High-Value Biologics – From Vaccines to Therapeutic Proteins, Lessons of the Past Applied Today and Tomorrow
Florian M. Wurm, Dr. rer. nat., Honorary Professor, Swiss Federal Institute of Technology Lausanne (EPFL); CEO,
ExcellGene SA
The talk reviews the dramatic history of our industry -- 100 years -- towards high-value biologicals whose manufacturing requires a huge range of knowledge of fundamental and applied sciences in fields as diverse as molecular biology to fluid dynamics
of water and gas. Reflecting and analysing the involved technologies in production provide excellent leads and opportunities for the future. CHO cells in bioreactors will be a central part of this discussion.
09:20 The Effect of Vector Design on the Expression of Bispecific Antibodies
Jason Saunders, MSc, Senior Scientist, Biologics Expression and Technology, Merck & Co., Inc.
09:50 Complementary Approaches for Protein Production in Insect and Mammalian Cell Lines
Konrad Büssow, Ph.D., Research Scientist, Structure and Function of Proteins, Helmholtz Centre for Infection
Research
The benefits of the baculovirus system for the production of intracellular and secreted proteins are well known. The human HEK293 cell line represents a valuable alternative, especially for secreted proteins, and allows for virus-free transient transfection
with plasmids. Recently, virus-free, plasmid-based transfection of insect cells has been optimised considerably. This system represents a useful alternative that combines advantages of the baculovirus system and transient transfection of HEK293 cells.
Examples of proteins produced by transient transfection of HiFive insect cells will be presented, including intracellular and secreted production.
10:20 Strep-Tactin XT - A Superior Next-Generation System for Purification of Proteins, Isolation of Cells & Assay Development
Dennis Niermeier, M.Sc., Scientist, IBA GmbH
The new third-generation Strep-tag® system is based on recently engineered Strep-Tactin®XT and Twin-Strep-tag®. Due to the affinity improved but still reversible binding of Strep-Tactin®XT to Twin-Strep-tag® in the low pM range,
the system is superior to other affinity purification systems and now also suitable for assay development.
10:35 Presentation to be Announced
10:50 Coffee Break in the Exhibit Hall with Poster Viewing
11:30 Engineering the Baculovirus Genome to Improve Protein Production
Dominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Laboratory
for Cancer Research, Leidos Biomedical Research, Inc.
The use of insect cell expression systems for the production of pharmaceutically relevant protein targets has dramatically increased over the last few years, with several products already approved by major regulatory agencies. In our laboratory, this
system has been vital for production of post-translationally modified RAS proteins essential for cancer drug discovery. In using this system, we have developed a number of process and technology improvements which permit increased protein yield,
protein quality, and virus stability. We discuss in detail the enhancements to the system and how they have been applied to high-level production of clinically relevant proteins, and examine ways in which synthetic biology and genome engineering
can further enhance the utility of this system.
12:00 SINEUPs: A New Class of Antisense Long Non-Coding RNAs that Specifically Activate Translation of Targeted Proteins
Silvia Zucchelli, Ph.D., Assistant Professor, Health Sciences, University of Eastern Piedmont; CSO,
TransSINE Technologies Inc.
SINEUPs represent a new functional class of natural and synthetic antisense long non-coding RNAs that upregulate translation of partially overlapping sense mRNAs through the activity of an inverted SINEB2 element. Given their modular structure, SINEUPs
can be designed to increase protein synthesis of potentially any gene of interest. We propose SINEUPs as reagents for molecular biology experiments, in protein manufacturing as well as in therapy of haploinsufficiencies.
12:30 Engineering of CHO Cell Lines for Enhanced Process Robustness
Pierre-Alain Girod, Ph.D., CSO, Selexis
High-quality production cell lines secreting maximal levels of recombinant proteins require stable integration of the recombinant DNA, elevated gene transcription, optimized secretion machinery to handle increased protein secretion and folding loads
and, ideally, being easily tracked during manufacturing. Using the data from our CHO-K1 genome and transcriptome, we have engineered our CHO-K1 to address these issues, particularly for difficult-to-express proteins, as well as to provide detailed
genomic analysis packages for manufacturing cell lines.
13:00 Luncheon Presentation: A
Stable Episomal Expression System to Streamline Mammalian Protein Production
Meelis Kadaja, Ph.D., MBA, Director, Business Development, Icosagen Cell Factory OÜ
We have found the way to stably maintain expression vectors in dividing mammalian cells as extrachromosomal units. This stable episomal expression system is scalable, and requires only 1ug of DNA to produce up to gram quantities of recombinant proteins
with low endotoxin levels in few weeks. Our system enables to generate production cell banks in 10 days, and is also used in antibody discovery to express and screen antibodies.
13:30 Session Break
14:00 Chairperson’s Remarks
Nicola Burgess-Brown, Ph.D., Principal Investigator, Biotechnology, Structural Genomics Consortium (SGC), University of Oxford
14:05 CHO Cell Energetics: Understanding the Powerhouse of the Cell through Mitochondrial Deep Sequencing
Paul S. Kelly, Ph.D., Senior Postdoctoral Researcher, School of Biotechnology, National Institute for Cellular
Biotechnology, Dublin City University
The predominant metabolic pathways, glycolysis and oxidative phosphorylation, have been linked to critical bioprocess relevant CHO cell phenotypes, growth and productivity. We have shown SEAP producing CHO cells engineered to be depleted of microRNA-23b
exhibited a 3-fold increase in product yield associated with elevated mitochondrial activity. Given CHO cells’ genetic instability, we sought to compare the mitochondrial genomic sequence of 22 CHO cell lines and provide a mitochondrial
genome reference sequence from the hamster.
14:35 Precision Control of Recombinant Gene Expression for CHO Cell Synthetic Biology
Adam Brown, Ph.D., Research Fellow, Department of Chemical and Biological Engineering, University of Sheffield
To successfully apply the core concepts underpinning synthetic biology to CHO cell engineering, we must develop practical and robust enabling technologies. Fundamentally, we will require the ability to precisely control the relative stoichiometry
of numerous functional components that are simultaneously introduced into the host cell factory. In order to enable this, we have developed a suite of complementary technologies that enable precise control of recombinant gene expression in
CHO cells. Using synthetic promoters as an exemplar, the function of these tools in the context of CHO cell engineering will be described.
15:05 Inactivation of GDP-Fucose Transporter in CHO Cells: A New Strategy for Producing Fucose-Free Biobetter Antibody Therapeutics
Zhiwei Song, Ph.D., Principal Scientist, Expression Engineering Group, Lead PI for GlycoSing Programme,
Bioprocessing Technology Institute, A*STAR
Removal of core fucose from IgG1 has been shown to significantly enhance its affinity to FcgRIII and thereby dramatically improves its antibody-dependent cellular cytotoxicity (ADCC). To address these industry-specific needs, we have inactivated
the GDP-fucose transporter in CHO cells. The DHFR and GS genes have also been inactivated in these cells as selection markers. Using these cells, stable lines have been developed to produce fucose-free rituximab and fucose-free obinutuzumab
(Gazyva).
15:35 Refreshment Break in the Exhibit Hall with Poster Viewing
16:15 A Big Step Forward for Next-Generation CHO Clone Characterization and Selection
Oliver Popp, Dr. rer. nat., Senior Scientist, Pharma Research and Early Development, Large Molecule Research,
Roche Innovation Center Munich, Roche Diagnostics GmbH
In-depth characterization of high-producer cell lines and bioprocesses is vital to ensure robust and consistent production of recombinant therapeutic proteins in high quantity and quality for clinical applications. For that, we established
a novel hybrid approach for supporting comprehensive characterization of metabolic CHO clone performance. The proposed approach also provides a mechanistic link between observed clone phenotype, process setup, and feeding regimes, and
thereby offers concrete starting points for subsequent process optimization.
16:45 FEATURED PRESENTATION: Engineering the CHO Cell
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Bjørn
Voldborg, MSc, Director, CHO Cell Line Development, Novo Nordisk Foundation Center for Biosustainability (CFB), DTU Biosustain, Technical University of Denmark
Using high-throughput (HT) technologies, the CHO Cell Line Engineering project at the Center for Biosustainability is genetically modifying CHO cells based on experimental and in silico generated data, to engineer CHO cell lines optimised
for the production of therapeutic proteins. The HT cell line engineering pipeline as well as examples of the engineered improved cell lines will be described.
17:15 Cmax: A Heterotrophic Cell Platform for Improved Biologics Production
Nicky C. Caiazza, Ph.D., Principal Scientist, Metabolic Engineering and Synthetic Biology, Heterotrophic Cell Systems, Synthetic Genomics Inc.
We have developed a robustly fermentative heterotrophic system for efficient end-to-end biologics discovery, development and manufacturing. This platform has intrinsic advantages for biologics production and a suite of synthetic biology tools
that has enabled high productivity (>1 g/l/d) mAb production with beneficial attributes for downstream processing. Here we show that these organisms can be engineered to secrete genuine, functional monoclonal antibody with favorable
characteristics related to protein production and quality.
17:45 Problem-Solving Breakout Discussions
Common Issues with Transient Protein Production
Moderators:
Richard Altman, MS, Scientist, Protein Technologies, Amgen, Inc.
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
Engineering Cell Factories for Protein Production
Moderator:
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Bjørn
Voldborg, MSc, Director, CHO Cell Line Development, Novo Nordisk Foundation Center for Biosustainability (CFB), DTU Biosustain, Technical University of Denmark
- Which production hosts do we engineer: microbial, insect and/or mammalian?
- How do we identify engineering targets: modeling, omics and/or literature?
- Which traits are we improving: product yield, product quality, host phenotype, process?
- Which proteins do we produce: antibodies, hard-to-produce proteins, other?
18:15 Networking Reception in the Exhibit Hall with Poster Viewing
19:15 End of Day
Day 1 | Day 2 | Speaker Biographies | Download Brochure
THURSDAY 3 NOVEMBER
08:00 Registration and Morning Coffee
08:30 Chairperson’s Remarks
Henry C. Chiou, Ph.D., Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific
08:35 Expression of IgG in Genetically Engineered E. coli
Na Ke, Ph.D., Research Scientist, Protein Expression and Modification Division, New England Biolabs
We have expressed for the first time in the cytoplasm of E. coli full-length human, rabbit and mouse antibodies, along with chimeric versions, including the commercial blockbuster Humira. The expression is further improved by
the co-expression of a series of protein-folding helpers. Microbial expression of IgG offers an expanded potential for rapid screening, engineering and expression antibodies.
09:05 Meeting Demand: Optimisation of Our Transient Expression Platforms to Increase Throughput and Titre
Christina Gordon, Scientist, New Meds, UCB Pharma
Development of antibody therapeutics, from early stage research to preclinical and clinical development, requires ever-increasing amounts of reagents. To meet the challenge of furnishing a diverse and full pipeline, we utilise several
different transient platforms. Through continuous optimisation, streamlining and automation of component parts of our panel of platforms (utilising both HEK293 and CHO host cells with a variety of transfection methods), we now have
the capability to produce microgram-to-gram quantities of panels of purified antibodies and antibody fragments in as few as four weeks from receipt of plasmid DNA.
09:35 Server of Many Masters: The Challenges of a Slim Protein Expression Core Facility
Tsafi Danieli, Ph.D., Director, BioGiv Excubator & Head, Protein Expression Facility, Wolfson
Centre for Applied Structural Biology, Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem
Running a small core facility that serves multiple academic groups with a wide range of disciplines is extremely challenging. Trying to do it with one technician, two postdocs and no automated platforms sounds like mission impossible.
However, it is all about the surrounding! An academic environment and supporting ecosystem allowed our facility to turn the challenge into a new model that successfully accommodates not only academic groups but also early stage biotech
startups.
10:05
Application of Streamlined Processes to Improve Process Productivity and Throughput
Naz Dadehbeigi, Pharm.D., Ph.D., Principal Scientist, Fujifilm Diosynth Biotechnologies
10:35 Coffee Break in the Exhibit Hall with Poster Viewing
11:15 Efficient Production of Recombinant Human Monoclonal Antibodies from Single B Cells
Hugo Mouquet, Ph.D., Head, Laboratory of Humoral Response to Pathogens, Immunology, Institut Pasteur
The molecular dissection of anti-pathogen B-cell responses using modern technologies to efficiently generate and characterize antigen-specific human monoclonal antibodies has allowed breakthrough discoveries in antiviral responses to viruses
such as Influenza virus and HIV-1. These recombinant antibodies represent unique “fingerprints” for each B-cell clone and when characterized at a molecular and functional level, provide crucial information to help developing
therapeutic and vaccine strategies against a given pathogen.
11:45 FEATURED PRESENTATION: Expressing Challenging Proteins at the SGC
Nicola Burgess-Brown, Ph.D., Principal Investigator, Biotechnology,
Structural Genomics Consortium (SGC), University of Oxford
The SGC has solved >1700 human protein structures and 6 novel integral membrane proteins. Although we have significantly contributed to structural biology and protein production for functional studies, there are many highly desired
targets including protein-protein complexes and known drug targets that remain a challenge to obtain. We present our established expression systems using E. coli and BEVS, and new processes (BacMam and mutagenesis) to
generate the most difficult-to-produce proteins.
12:15 Enjoy Lunch on Your Own
13:00 Dessert Break in the Exhibit Hall with Poster Viewing
13:30 End of Optimising Expression Platforms
Day 1 | Day 2 | Speaker Biographies | Download Brochure