Cambridge Healthtech Institute’s 7th Annual
Optimizing Cell Line Development
Enhancing Expression
August 6-7, 2015
Part of CHI's 7th Annual The Bioprocessing Summit
August 3-7, 2015 | Westin Copley Place Hotel | Boston, Massachusetts
The “Optimizing Cell Line Development” meeting features experts sharing their case studies and anecdotes that illustrate the strategies they use to reach enhanced expression and lower costs. Their research and experiences provide meaningful
insights into how to best select cell lines and hosts, and develop high-producers into late-stage production. Gaining a greater understanding of cells through sequencing and the omics sciences will also be addressed, including the CHO genome research,
metabolomics, assays, and QPCR. In addition, challenges for introducing new technologies will be discussed, along with an overview of industrial trends and regulatory perspectives.
Thursday, August 6
1:55 pm Chairperson’s Remarks
Christina Alves, Ph.D., Scientist, Cell Culture Development, Biogen Idec, Inc.
2:00 OPENING KEYNOTE PRESENTATION:
The Future Looks Great…but What Is It?
Alan Dickson, Ph.D., Professor and Director, Centre of Excellence in Biopharmaceuticals (COEBP), University
of Manchester
Use of understanding developed through systems biology, coupled to the application of synthetic biology, will generate an exciting and brave manufacturing world in which novel CHO cell variants will be matched to expression of engineered unique biopharmaceutical
entities … perhaps? I will give a personalized review of the reality of progress to date and how adoption of developing technologies has the potential to make a real difference.
2:45 Upstream Process Optimization, Automation and Collaboration: An Accelerated Path to the Clinic
Pamela Pegman, Ph.D., Senior Principal Scientist, Cell Line Development, Pfizer, Inc.
Optimizing cell line development activities has the potential to reduce the time and resources required for clinical entry. This presentation will summarize a combination of approaches to reduce cell line development timelines including creative timing
of upstream activities through collaboration and harmonization with discovery partners, introduction of automation, and the use of business practice modeling to predict the best outcomes for key technology introduction.
3:15 SELECTED POSTER PRESENTATION
A29 Bioprocess for Testing Engineered CHO Cell Lines Using Single-Use Bioreactors
Marianne Decker, Ph.D., Special Consultant, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
3:45
Using Combinatorial Genome Editing to Improve Production in CHO Cells
Brian Burke, Ph.D., Business Development Director, Products, Horizon Discovery
Horizon have started a combinatorial gene editing program to improve CHO cell line performance. Together with the development of a high throughput reporter system we will generate a range of cell lines with different production traits to meet the
future needs of biologics manufacture and develop next-generation production systems.
4:00 Ice Cream Social in the Exhibit Hall with Poster Viewing
4:45 Engineering the Baculovirus Genome for Better Protein Production from Insect Cells
Dominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Lab for
Cancer Research
Protein production from insect cells using the baculovirus expression system has proven very useful for the generation of high-quality heterologous proteins when other systems have failed. However, insect cell protein production differs from mammalian
expression due to differences in post-translational modification pathways. In order to overcome some of these differences, genome engineering of the baculovirus can be used to “humanize” insect cell protein production. We describe
methods for facile modification of the baculovirus genome to both add new pathways for post-translation modification of proteins and also to enhance the stability of baculovirus constructs for protein production.
5:15 Optimizing Insect Cell Lines for Production of Structurally Uniform Glycoproteins
Donald Jarvis, Ph.D., Professor, Molecular Biology, University of Wyoming
My presentation will cover previous, as well as more recent and ongoing efforts to produce insect cells with humanized glycoprotein processing pathways. Data will show recent efforts have yielded insect cell lines optimized for human-type protein
glycosylation. Furthermore, data will show those cell lines can produce structurally homogenous glycoproteins.
5:45 End of Day
6:30-9:00 Recommended Dinner Short Course
Transient Protein Production in Mammalian Cells
Friday, August 7
8:00 am Registration and Morning Coffee
8:25 Chairperson’s Remarks
Alan Dickson, Ph.D., Professor and Director, Centre of Excellence in Biopharmaceuticals (COEBP), University of Manchester
8:30 FEATURED PRESENTATION:
An Inducible System for the Rapid Generation of CHO Pools and Stable Clones
Yves
Durocher, Ph.D., Research Officer, Human Health Therapeutics Portfolio, Biologics & Biomanufacturing Program, National Research Council Canada
Using the cumate-inducible promoter, we developed a CHO platform for the rapid generation of stable pools in less than 3 weeks post-transfection. The pools are stable over time and can be used to isolate stable CHO clones showing high productivities.
We will provide examples of pools generated for the production of monoclonal antibodies, secreted and membrane proteins. This platform is a useful and cost-effective addition to the large-scale CHO transfection platform for the rapid production
of recombinant proteins.
9:00 Screening of CHO Cell Clones by Mass Balance of Amino Acids
Wen Wang, Ph.D., Postdoctoral Associate, Chemical Engineering, Massachusetts Institute of Technology (MIT)
Post-transfection clone screening is a crucial step in mammalian cell line development for production of therapeutic proteins. Ideally, the screening should be non-invasive, economical, and effective. We report a fundamentally new method for predicting
mAb titer in a Chinese hamster ovary (CHO) cell culture system based on mass balance of two essential amino acids. This study provides an alternative way to screen high therapeutic protein producers in a low-cost manner.
9:30 Stable Glycoengineering of CHO Cells
Claus Kristensen, Ph.D., Associate Professor, Copenhagen Center for Glycomics (CCG),
University of Copenhagen
Recent advances in precise gene editing technologies such as ZFNs, TALENs and CRISPR/Cas9 systems have enabled stable engineering of mammalian cells to produce well-defined N and O-glycans. In this lecture we will present state-of-the-art glycoengineering
in CHO to produce more homogeneous glycans and options for novel designed glycan structures.
10:00 Coffee Break
10:45 Developing Systems Biology Models Based on Whole-Genome Sequencing of CHO to Guide Cell Line Engineering
Nathan Lewis, Ph.D., Assistant Professor, Systems Biology Research Group, University of California,
San Diego
Our recent whole-genome sequencing efforts for CHO have enabled the construction of systems biology models of metabolism, protein secretion, and glycosylation. We now use these for detailed analysis of -omics data from CHO to deepen our understanding
of differences between host cell lines and to guide our cell line engineering efforts. These efforts aim to enhance cell growth, tailor protein modifications, and improve protein secretion, in order to better control biotherapeutic critical
quality attributes.
11:15 Utilizing High-Throughput Bioanalytics for Early Detection of Product Impurities
Christina Alves, Ph.D., Scientist, Cell Culture Development, Biogen Idec, Inc.
Improvements in the throughput and efficiency of analytical techniques have enabled their use earlier on in the cell line development process. This in turn provides useful information on a large number of clones at a very early stage. In this
case study we demonstrate how the use of these techniques allowed early detection of product impurities which in turn prevented delays in clinical timelines and the need for additional resources.
11:45 High-Throughput Product Quality Assays for Cell Line and Process Development
Shashi Prajapati, Ph.D., Senior Scientist, High-Throughput Analytical Group, Cell Culture Development, Biogen Idec, Inc.
Cell line and process development play a major role in producing therapeutic proteins with high productivity and appropriate product quality attributes. To support this development, analyses of large number of samples generated from thousands
of clones and process optimization are critical. Analyzing large number of samples has been a bottleneck in biotech industries because conventional analytical assays are low-throughput. Here we present various high-throughput (HTP) analytical
platforms to facilitate rapid and parallel analyses of product quantity and quality using 96-well plate formats. These platforms include HTP protein quantitation followed by HTP protein purification and product quality analyses. With these
analytical capabilities, we can assess product quality in the early stage of clone screening, as well as expedite the cell line and process development.
12:15 pm Late Breaking Presentation
12:30 Luncheon Presentation: Preventing Virus Contamination of Bioreactors with a Virus Barrier Filter
Christina Carbrello, Ph.D., Senior Scientist, Process Solutions, EMD Millipore
Cell culture media is typically sterile filtered to remove bacteria and mycoplasma, however many bioreactors remain unprotected from viral contamination. Ideally, a filter could be used to remove viruses, but filtration is traditionally perceived
as being unsuitable for the upstream processes. A new virus filter has been evaluated and demonstrated high levels of virus, bacteria and mycoplasma removal while providing high flow and capacity in chemically defined media with little
to no impact on cell culture performance.
1:15 Session Break
1:25 Chairperson’s Remarks
Pamela Pegman, Ph.D., Senior Principal Scientist, Cell Line Development, Pfizer, Inc.
1:30 Development of Synthetic Biology Tools to More Predictably Clone, Express and Select Active Biologics in Cell Line Development
Ian Fotheringham, Ph.D., Managing Director, Ingenza Ltd.
In this talk, I will describe how Ingenza has developed and deployed various synthetic biology tools to enable us to more predictably clone, express and select active proteins during cell line development. I will use real-world examples of solving customer problems to illustrate the utility of our approach. These tools include protein engineering to address poor response kinetics during the control of gene expression, the development of synthetic “landing pads” to optimize the genomic operating environment, and the use of genome editing and RNA trafficking systems.
2:00 Enabling Evaluation of Polysaccharide Size and Content for DOE Studies in Early Process Phases
Erwin Swennen, Ph.D., Manager, Cell Line and Process Analytics, Bacterial Drug Substance Development, Novartis Vaccines
This talk presents the development of an innovative RP-SE(GPC) analytical method to monitor both critical quality attributes of GBS capsular PS during all phases of the production process. This method enabled DOE studies also in early
process phases with complex matrix, and contains a practical approach on how industrial R&D is trying to keep up with the constant increase in quality requirements by regulatory agencies (QbD, DoE in early phases). The method
represents a new out-of-the-box application of technology.
2:30 Development of Human Cell Systems for Evaluating the Responses of Drugs Directed at Correcting the Function of Cellular Proteins
Dieter Gruenert, Ph.D., Professor, Otolaryngology-Head and Neck Surgery, University
of California, San Francisco
Our lab has been a leader in the development of these cell systems that have been used in academic and industry settings nationally and internationally. We have developed human airway epithelial and iPS cell systems to evaluate and
screen the responses to drugs aimed at correcting the function of proteins involved in airway diseases such as cystic fibrosis.
3:00 GlycoExpress: A Toolbox for the High Yield Production of Glycooptimized Fully Human Biopharmaceuticals in Perfusion Bioreactors at Different Scales
Rainer Stahn, Ph.D., Head, Bioprocess Development, Glycotope GmbH
GlycoExpress cells producing mAb are cultivated with perfusion bioreactor systems applying different cell retention mechanisms such as centrifugation (centritech) or alternating tangential flow (ATF) filtration at different scales.
Growth, product yield and product quality, especially glycosylation, are evaluated during the cultivation. Perfusions culture helps to keep cells in the optimal growing and production phase over the production process which leads
to highly stable product quality allowing a flexible duration of the run in one batch size.
3:30 Close of Conference