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Optimizing Mammalian Cell Lines - Day 2


Optimizing Mammalian Cell Lines 


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Thursday, August 25

7:30 am Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee

 

OPTIMIZING PROPERTIES 

8:25 Chairperson's Remarks

Dieter C. Gruenert, Ph.D., Professor, Department of Otolaryngology, Head and Neck Surgery, Department of Laboratory Medicine, University of California, San Francisco

8:30 Analysis of the RNAi Pathway for Improved siRNA Design

S. Patrick WaltonS. Patrick Walton, Ph.D., Associate Professor, Chemical Engineering & Materials Science, Michigan State University - Biography 

RNA interference (RNAi) provides a powerful means for down regulating the expression of a specific gene in mammalian cells. However, selection of the most active short, interfering RNA (siRNA) against a target is generally accomplished by guided trial-and-error. We will discuss our results on the interactions of siRNAs with other molecular components of the RNAi pathway and how these interactions inform siRNA design and selection.

9:00 Monitor and Control of Glycosylation in Mammalian Cell Bioprocesses

Michael ButlerMichael Butler, Ph.D., Distinguished Professor, Animal Cell Technology, Microbiology, University of Manitoba

Critical culture parameters control the glycosylation profile of proteins secreted from mammalian cells in culture. The parameters may be associated with the host cell line, the culture media, the mode of culture or 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. These parameters will be discussed in the context of bioprocesses for the production of highly efficacious biopharmaceuticals with consistent structural profiles.

9:30 Prospects for Generating Cell-Type Specific Immortalized Cells

Dieter GruenertDieter C. Gruenert, Ph.D., Professor, Department of Otolaryngology, Head and Neck Surgery, Department of Laboratory Medicine, University of California, San Francisco - Biography 

Blastocyst and Induced Pulripotent Stem Cells Derived from Sv40-Transformed Tracheal Epithelial Cells by Somatic Cell Nuclear Transfer or Reprogramming.  The prospect of using cell models to study disease pathology and develop new therapies has been significantly enhanced by stem cell technology. In particular, somatic cell nuclear transfer (SCNT) and the generation of induced pluripotent stem cells (iPSCs) has opened the door to new opportunities for developing cell-type specific immortalized cell systems.  In this context, we have generated blastocysts through SCNT of SV40-transformed rabbit tracheal epithelial cell nuclei and iPSCs through cellular reprogramming for differentiation into cell-type specific immortalized cells.  These cells can now be used for evaluating therapeutic interventions in multiple tissue types in the same genetic background and have the potential for developing and optimizing patient specific therapies.

10:00 Networking Coffee Break

 

OPTIMIZING CLONES 

10:30 Differential Growth, Productivity, Metabolism and Responses to Trophic Factors in CHO Cell Clones

Susan SharfsteinSusan Sharfstein, Ph.D., Associate Professor, Nanobioscience, College of Nanoscale Science and Engineering, University at Albany - Biography 

To evaluate the range of possible responses to changes in process conditions, the growth, metabolism, and productivity of five Chinese hamster ovary (CHO) clones were explored in response to stimulation with insulin (5 mg/L) and LONG®R3IGF-I (20 µg/L or 100 µg/L). All five clones were derived from the same parental CHO cell line (DG44) and produced the same recombinant monoclonal antibody, with varying specific productivities. Overall product titers were affected by variations in both integrated viable cell density (IVCD) and specific productivity. Nutrient uptake and metabolite generation patterns varied strongly between clones and much less with culture conditions. These results point to the need for careful clonal analysis when selecting clones, particularly for platform processes where media and culture conditions are predetermined.

11:00 Fluorescent Labelling in Semi-Solid Medium to Identify High-Expressing Clones for Recombinant Protein Production

Renald GilbertRénald Gilbert, Ph.D., Research Officer, Biotechnology Research Institute, National Research Council Canada - Biography 

We have optimized a method to identify those valuable but rare supersecretors. It consists of directly labelling small colonies of CHO cells with a fluorescent antibody against the secreted gene product in semi-solid medium. Because a correlation exists between the intensity of the fluorescent signal and the level of secreted protein, the few supersecretors within a pool of transfected cells can be easily identified and isolated using a micromanipulator or an automated cell picker. This method reduces considerably the work load necessary to generate stable high-expressing cell lines, because only a small number of clones need to be subsequently analyzed for protein production and stability.
 

11:30pm End of The Bioprocessing Summit



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