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Optimizing Cell Culture Technology conference - Day 2

Optimizing Cell Culture Technology 

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8:00 am Registration and Morning Coffee



8:25 Chairperson’s Remarks

Yao-Ming Huang, Ph.D., Principal Engineer, Biogen Idec, Inc.


Novel Approaches for the Prediction and Isolation of Highly Productive Recombinant CHO Cell Lines

Mark-SmalesMark Smales, Ph.D., Professor, Biotechnology, School of Biosciences and Centre for Molecular Processing, University of Kent - Biography 

The ability to generate high producing recombinant mammalian cell lines has been greatly enhanced in recent years through the development of approaches such as high-throughput screening technologies. Despite this, the ability to predict the phenotypic behavior of specific cell lines in the bioreactor early in the cell line development process remains challenging. We have developed new screening and predictive performance approaches that allow the prediction of a cell line’s performance at the bioreactor scale early in the cell line construction process. Here this approach and its application to cell line construction will be discussed.

9:00 Cell Culture Platform Application in an Early-Stage mAb Project

Yao-Ming-HuangYao-Ming Huang, Ph.D., Principal Engineer, Biogen Idec, Inc.- Biography 

9:30 Towards Dynamic Metabolic Flux Analysis in CHO Cell Cultures

Maciek-AntoniewiczMaciek R. Antoniewicz, Ph.D., Assistant Professor and DuPont Young Professor, Chemical & Biomolecular Engineering, University of Delaware - Biography 

The metabolism of Chinese hamster ovary (CHO) cells changes dramatically during a fed-batch culture as the cells adapt to a changing environment and transition from an exponential growth phase to stationary phase. In this talk, I will highlight progress on 13C-metabolic flux analysis in CHO cells and discuss novel techniques for dynamic metabolic flux analysis. Application of these new tools may allow identification of intracellular metabolic bottlenecks at specific stages in CHO cell cultures and eventually lead to novel strategies for improving CHO cell metabolism and optimizing biopharmaceutical process performance.

TAP Biosystems10:00 Characterization of a High-Throughput Micro Bioreactor: Process Control, Consistency and Comparability in CHO Clone Ranking and Process Optimization Studies

Tim Ward, Director, Strategic Product Marketing, TAP Biosystems

The ambrTM system combines the benefits of single use labware and automated liquid handling to provide a high-throughput microscale bioreactor system with capability for both batch and fed-batch cell cultures. This talk introduces the ambr, its potential in bioprocess development and, through discussion of field data gathered to date, demonstrates its effectiveness in CHO cell culture applications: clone screening, process optimization, and design of experiments.

10:15 Coffee Break in the Exhibit Hall with Poster Viewing



11:00 Dynamic Transcriptome Profiling for Cell Culture Process Comparisons

Karthik P. Jayapal, Ph.D., Staff Scientist, Cell Culture Development, Global Biological Development, Bayer HealthCare Pharmaceuticals

Existing literature on genomics for mammalian cell culture pertains primarily to non-producing cells or those in exponential growth phase which may not have direct relevance to commercial manifacturing. In this study, we employed DNA microarrays to analyze the temporal transcriptome dynamics of commercial recombinant protein-producing cells in two processes – (1) a long-term high-flow non-mAb perfusion process and (2) a ~2 week mAb fed-batch process. In the former case, despite the presumable steady-state nature of the perfusion process, we identified certain subtle changes in the cell’s transcriptome which correlated with cell age while, at the same time, demonstrating the utility of genomics technologies for scale-down model validation. In the latter example, we show a preliminary investigation of lactate production and consumption phenotypes to identify clusters of genes correlated with lactate production.

11:30 Mammalian Cell Fluid Mechanics and Scale-Up Considerations

Jeffrey J. Chalmers, Ph.D., Professor, Chemical & Biomolecular Engineering, Ohio State University

Suspension animal cell culture is now routinely scaled up to bioreactors on the order of 10,000 liters, and greater, to meet commercial demand. However, the concern of the “shear sensitivity” of animal cells still remains, not only within the bioreactor, but also in the downstream processing. The perception of “shear sensitivity” has historically put an arbitrary upper limit on agitation and aeration in bioreactor operation; however, as cell densities and productivities continue to increase, mass transfer requirements can exceed those imposed by these arbitrary low limits. This presentation will mainly focus on publications from both academia and industry regarding the effect of hydrodynamic forces on industrially relevant animal cells, and on general observations with respect to scale-up.

12:00 pm Accelerating Bioprocess Optimization through the Use of Next-Generation Genomics Technologies

Len-van-ZylLen van Zyl, Ph.D., Research Assistant Professor, College of Natural Resources, North Carolina State University; CEO, ArrayXpress, Inc. - Biography 

In this presentation, we explain the financial and scientific benefits of incorporating Next-Generation Genomics (NGG) technologies into standard practices for research and development of therapeutic biologics and their manufacturing development and optimization. We will give experience-based examples of how a systems biology approach improved our understanding of a production organism’s biology, and provided detailed insight of how the cell’s genetic machinery responded to changes in Critical Process Parameters. The results directly contributed to improved product yields, product quality, and overall process stability.

Pall_LifeSciences 12:30 Luncheon Presentation
A New Bench Scale Single Use Bioreactor System
Charles Golightly, Ph.D., Director of Marketing, Pall Life Sciences
Development of a new bench scale “rocker style” single use bioreactor will be described, with discussion of physical parameters for design space definition and performance measurement; with comparison to industry standard systems.    Presentation will conclude with results from CHO batch culture trials for design validation, showing significant increases in achievable cell densities and cell productivity. 


1:55 Chairperson’s Remarks

Jeffrey J. Chalmers, Ph.D., Professor, Chemical & Biomolecular Engineering, Ohio State University

2:00 Assessing and Controlling the Raw Material Variability in Mammalian Cell Culture

Seongkyu Yoon, Ph.D., Assistant Professor, Chemical Engineering & Director, BioManufacturing Center, University of Massachusetts, Lowell

Biologics manufacturers are facing challenges due to lot-to-lot variations of critical raw material, lack of appropriate measurements of intermediate process parameters, and even inappropriate analytical test methods of final product quality attributes. The presentation will illustrate how to characterize raw material variations and its impact in cell culture processes. Raw materials and media will be analyzed for lot-to-lot variation characterization using analytical spectroscopies. The collected data will be used to develop multivariate regression models between raw material and cell-culture performance for prediction of viable cell density and cell viability. These models can be used for subsequent batches to compensate raw material variabilty and accurately predict the cell-culture performances.

2:30 Control of Glycosylation Profiles in Cell Bioprocesses

Michael-ButlerMichael Butler, Ph.D., Professor, Microbiology, University of Manitoba - Biography 

The glycosylation profile of proteins secreted from mammalian cells in culture is dependent upon critical parameters associated with the host cell line, the culture media, the mode of culture and the specific protein synthesized. It is important to control these parameters in an industrial bioprocess to ensure consistency of the final product and maximum bioactivity. The critical culture parameters will be discussed in the context of bioprocesses in which fine control is necessary to synthesize glycoproteins for the production of highly efficacious biopharmaceuticals with consistent structural profiles.

3:00 Permittivity Measurements in Cell Culture Processes

Sven-AnsorgeSven Ansorge, Ph.D., Development Scientist I, Alexion Montréal Corp. - Biography 

In routine cell culture operations, process characterization and supervision relies primarily on offline sampling and analysis resulting in extended response times to process events and variations. Further efforts are thus needed to implement real time monitoring tools that allow for advanced and rationalized process control strategies. We provide here insights into the recent progress made concerning the use of in situ permittivity-based sensors in cell culture processes. Published and unpublished findings from several groups including work on CHO, insect (Sf-9) and HEK293 cell lines are presented and discussed. It is demonstrated that this technology allows tracking of the biovolume content (i.e., cell growth), the metabolic activity and, in particular in the case of viral productions, product release.

3:30 Refreshment Break in the Exhibit Hall with Poster Viewing



4:15 Taking Cell Culture to the Third Dimension

Chi-ZhangChi Zhang, Ph.D., Fellow, Division of Nanobiotechnology, School of Biotechnology, Royal Institute of Technology (KTH) - Biography 

Highly functional in vitro cultured cells are of great usefulness in various applications such as cell-based testing, constructing large tissues and pathological research. The key to culturing functional cells in vitro is to recapitulate an in vivo-like cellular microenvironment which allows extensive mechanical support as well as cell-cell, cell-matrix and cell-soluble factor interactions. The development of 3D cell culture devices has made possible the controlling of cellular microenvironments, phenotypes and behaviors under novel experimentations. Microfluidic systems are believed to be advantageous over other systems in terms of a more controllable microenvironment, however, few attempts to engineer a soluble microenvironment for extensive cell-soluble factor interactions within these microfluidic systems are reported. Hence, there is a great need to develop an in vitro model with well-controlled soluble microenvironment to primarily supplement in vivo animal models, thus reducing the cost and ethical issues surrounding animal experimentation.

4:45 A Scaffoldomics Approach to Optimizing Synthetic 3D Cell Niches

Carl-SimonCarl G. Simon, Jr., Ph.D., Biologist, Polymers Division, Biomaterials Group, National Institute of Standards & Technology (NIST) - Biography 

We have developed scaffold library approaches to systematically screen the effect of scaffold properties on cell function in 3D. Our most recent work has focused on the effect of 3D scaffold structure on the morphology and differentiation of primary human bone marrow stromal cells (hBMSCs). We have found that some scaffold structures promoted hBMSC differentiation while others enhanced proliferation. Microarray screening revealed that each scaffold structure induced a unique and reproducible gene expression signature, indicating that hBMSCs had a precise response to each architecture. Hierarchical cluster analysis showed that treatments sorted by scaffold structure and not by composition, suggesting that structure was more important than chemistry. The effects of scaffold structure appeared to be mediated by cell shape, where substrates that induced differentiation also induced a similar hBMSC morphology.

5:15 End of Conference

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