Cambridge Healthtech Institute’s Third Annual
CHO Cell Lines
Enhancing Expression, Performance and Process
January 11-12, 2017 | Hilton San Diego Bayfront | San Diego, CA
CHO cells’ rapid rise in production prominence is due to their adaptability to various culture conditions, gene plasticity, and ability in proper folding, posttranslational modifications, and glycosylation of desired proteins. Thus, advances
in CHO cell lines and culture continue to significantly improve biotherapeutic production. This achievement is due to progress in engineering stable and transient cell lines, enhancing cell culture conditions and performance, as well as optimizing
process development. When all are accomplished, higher-production titers and better product quality result. The CHO Cell Lines conference gathers cell line engineers, cell culture specialists and bioprocess development managers to explore the
latest data, tools and strategies for improving protein expression, production, and product quality.
WEDNESDAY, JANUARY 11
1:00 pm Conference Registration
2:00 Chairperson’s Remarks
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
2:05 A Predictable, Plug-and-Play Cell Culture Platform Process
James Lambropoulos, MS, Engineer III, Cell Culture Development, Biogen
We have implemented a defined workflow for early stage clinical cell line development. A meta-analysis of several monoclonal antibody products, in multiple host cell lines, demonstrates predictable trends and correlations amongst growth, metabolite,
productivity, and quality attributes. This analysis confirms that our development platform is a robust and reliable workflow for generating representative, high-quality protein material for clinical use, and offers interesting avenues for further
refinement of the process.
2:35 In the Pursuit of High Producers: Use of the Sony SH800 Cell Sorter for the Selection of Cell Lines with Superior Productivity Characteristics
Nadia Amharref, Ph.D., Scientist, Cell Line Development, Vaccine Research Center, National Institute of Allergy
and Infectious Diseases, NIH
Shortening the process of developing and isolating high-producing cell lines remains a challenge. Our study focuses on the development of a simple and rapid method using Fluorescence-Activated Cell Sorting (FACS) for the screening of high-producing
recombinant CHO cell lines. In this method, flow cytometry was partnered with a reporter protein for rapid, early stage identification of clones producing high levels of a therapeutic protein. A cell surface protein not normally expressed on CHO
cells is co-expressed, as a reporter, with the therapeutic protein and detected using a fluorescently labeled antibody. The reporter protein’s expression level accurately predicts the relative expression level of the therapeutic protein
for each clone.
3:05 Fast Cell Line Development for CHO Clones with High-Yield Protein Production Using Euchromatin-Containing BAC Expression Vectors
Anton Bauer, Ph.D., COO, The Antibody Lab GmbH
Upon stable cell line generation, chromosomal integration site of the vector DNA has a major impact on transgene expression. By using chromosomal loci in BACs and random integration into host cell chromosomes, we developed stable high-yield production
cell lines at an unprecedented speed. We performed several case studies for CHO production clones, and we established for antibodies, and even difficult-to-express proteins, generation of production clones within three weeks from transfection.
3:35 Refreshment Break in the Exhibit Hall with Poster Viewing
4:30 A Method for Specifically Targeting Two Independent Genomic Integration Sites for Co-Expression of Genes in CHO Cells
Joop van den Heuvel, Ph.D., Research Group Leader, Recombinant Protein Expression, Helmholtz Centre for Infection Research
5:00 Engineering Protein Production Hosts
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for
Biosustainability, Technical University of Denmark
We are using the combined competencies of scientific groups working within the areas of metabolic modelling, glycobiology, cell line engineering, high-throughput methodology and genome editing tool development to design and engineer the next generation
of recombinant protein production hosts. The most recent results from high-throughput targeted genomic manipulations to engineer cells for tailored and homogenous glycosylation, increased productivity, improved product quality and more robust
bioprocesses will be presented.
6:20-7:20 Reception in the Exhibit Hall with Poster Viewing
7:20 Close of Day
THURSDAY, JANUARY 12
7:45 am Morning Coffee
8:15 Chairperson’s Opening Remarks
Sohye Kang, Ph.D., Senior Scientist, Process Development, Amgen
Keynote Presentations
8:20 Strategies to Enable High-Throughput Recombinant Protein Production in Mammalian Cells for Preclinical Studies
Athena Wong, Ph.D., Senior Scientist & Senior Group Leader, Early Stage Cell Culture, Genentech
Transient transfections in HEK293 and CHO cells are used to rapidly generate proteins for discovery research and early development studies. Here we present our approaches to express microgram to multigram amounts of protein in automated 96
deep well plates and bioreactors. To increase transfection productivity, we performed host cell engineering followed by process optimization. Results showed that modifying media components provides significant benefits towards increasing
yield and/or modulating product quality.
9:00 Today a Caterpillar, Tomorrow a Butterfly: The Transformation of Bioprocess Analytics
Beth Junker, Ph.D., Principal Consultant, BioProcess Advantage LLC
The volume and complexity of bioprocess data are increasing dramatically, requiring a transformation of bioprocess analytics to effectively extract meaningful information. Whether in clinical development or commercial manufacturing, tomorrow’s
focus is moving away from Big Data towards Smart Data approaches. Available tools such as Multivariate data analysis as well as other statistical methods overlay the necessary structure. This permits purposeful analytics which ultimately
reveal valuable insights into process and analytical understanding.
9:30 Engineering Process Performance by Predictive Insilico Solutions
Dirk Müller, Ph.D., Head, R&D Services, Insilico Biotechnology AG
Genome-based network models have been reconstructed for a number of the biotech industry’s workhorses and find increasing application for the quantification of bioprocess performance and for the evaluation of strain engineering strategies
in silico. The biotech industry, however, is still a far cry from the situation in other areas like electronics or automotive where each new product is designed and optimized employing computer simulations, visualization, and
data management solutions. Since costs for both genomic data and computing resources are decreasing rapidly, we also expect the bioeconomy to transform into a knowledge-based industry within the next decade. This transformation will lead
to (i) significantly reduced development timelines, (ii) increased process performance, and (iii) improved quality of life-saving therapeutics. Nevertheless, there is a rather long way to go because a full mechanistic understanding of
cellular systems is presently lacking and the software tools and algorithms to swiftly do the job for every bioprocess are yet to be developed. In this contribution, we present first steps we have taken in this direction with a technology
platform comprising both content and software. The platform supports efficient setup and curation of genome based models as well as integration of metabolite data and transcript levels. Dedicated software tools enable automated quantification
of process performance and derivation of predictive dynamic models. We illustrate the application of such predictive models to bioprocess optimization including media design and strain engineering and discuss chances and current challenges
of this approach.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing
FEATURED PRESENTATION
11:00 Genome-Scale Big Data and Modeling Approaches to Optimizing Protein Production in CHO Cells
Bernhard Palsson, Ph.D., Galletti Professor, Bioengineering; Principal Investigator,
Systems Biology Research Group, Bioengineering; Professor, Pediatrics, University of California, San Diego
Three technological drivers that advanced the development of microbial production strains are now in place for CHO cells. These are: 1) whole-genome sequences, 2) genome editing tools, and 3) genome-scale models. The last item is reliant on
big data analysis against a structured network reconstruction for metabolism and protein secretion. Network reconstructions are knowledge bases that formalize our knowledge of biochemistry, genetics, and genomics in CHO. These are called
BiGG k-bases. The history, development and status of this field is reviewed in this talk, along with a view of what the future may hold.
11:30 Session Break
12:15 pm Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:15 Ice Cream Break in the Exhibit Hall with Poster Viewing
2:00 Chairperson’s Remarks
Richard Altman, MS, Scientist, Protein Technologies, Amgen
2:05 Chromatin Function Modifying Elements in an Industrial Antibody Production Platform
Mark Ellis, Principal Scientist, Protein Expression and Purification, UCB Pharma
The isolation of stably transfected cell lines for the manufacture of biotherapeutic protein products can be an arduous process. This frequently involves transgene amplification and maintenance over many generations. We assessed four chromatin
function modifying elements for their ability to negate chromatin insertion site position effects and their ability to maintain antibody expression. Stability analysis demonstrated that the reduction in expression was mitigated in the
clones containing A2UCOE-augmented transgenes.
2:35 Cell Line Profiling to Improve Productivity and Product Quality
Sohye Kang, Ph.D., Senior Scientist, Process Development, Amgen
Despite the same host cell origin, recombinant production cell lines often display phenotypic variability in regard to growth rate, cell size and metabolic profiles. These intrinsic, cell line-specific variations can affect productivity and
product quality. Technological advances in -omics and bioinformatics tools are providing unprecedented opportunity to probe the molecular systems that underlie various cellular phenotypes influencing quantity and quality of therapeutic
protein production. Findings from these investigations allow opportunities for process improvement.
3:05 Accelerating Biotherapeutic Development through Simultaneous High-Titer, CHO Transient Expression & Generation of High-Yield Stable Cell Lines Using Scalable Transfection
James Brady, Ph.D., Director, Technical Applications, MaxCyte, Inc.
3:35 Refreshment Break in the Exhibit Hall with Poster Viewing
4:15 Product Quality and Protein Expression in CHO Cells
Pauline Smidt, Process Development Scientist, Just Biotherapeutics
I discuss the evaluation of growth, titer and product quality in production CHO cell lines under various process conditions.
4:45 Rapid Production of Recombinant Proteins in CHO Cells Using Large-Scale Transfection or Stable Pools
Yves Durocher, Ph.D., Section Head, Mammalian Cell Expression - NRC Human Health Therapeutics
Portfolio, National Research Council Canada
We describe our CHO transient expression platform for rapid production of recombinant proteins. For more difficult-to-express proteins or proteins needed in large quantities, we also developed an inducible CHO pool platform that allows generation
of stable and scalable pools expressing high levels of monoclonal antibodies in two weeks post-transfection. Fc glycans present on monoclonal antibodies produced by transient transfection or stable pools are compared. The pools can also
be used to derive stable and high-expressing CHO clones for manufacturing therapeutic candidates.
5:15 Toolbox for Cell Line Development – Next-Generation Cell Line Development Technologies
Holger Laux, Ph.D., Fellow, Integrated Biologics Profiling, Technical Development NBE, Novartis
Chinese hamster ovary (CHO) cells are the most widely used host for large-scale production of recombinant therapeutic proteins exhibiting high productivities in the gram-per-liter range. 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 evaluated novel vector elements and a new selection marker with a variety of antibody
projects resulting in an increase of titer. Furthermore, we have identified a key protein severely affecting the quality of non-antibody format therapeutic proteins. Subsequently the key protein was eliminated using Novartis propriety
CHO cell line via novel targeted gene disruption tools. This resulted in a superior CHO cell line. The combination of novel vector elements, novel cell lines and selection marker resulted in a significant increase of titers and improved
product quality.
5:45 Close of Conference