Cambridge Healthtech Institute’s 2nd Annual
Transient Protein Production
From Small-Scale to Large-Scale
January 22-23, 2015
Speed, limiting risk and protein quality are often cited as advantages of transient protein production (TPP). Transient expression achieves both fast process times and high yields because the gene of interest is simply transfected via a plasmid into the respective producer cells. In addition, compared to expression in lower-order organisms, recombinant protein expression in mammalian producer cells (CHO and HEK293) or alternative hosts offers significant advantages in protein quality and posttranslational modification. However, the rapidly increasing need for recombinant proteins necessitates further improvements in TPP technologies.
Cambridge Healthtech Institute’s Second Annual Transient Protein Production conference convenes protein expression specialists who share their experiences of the differences, tradeoffs, advantages and improvements in producing recombinant proteins in transient production systems.
Day 1 | Day 2 | Download Brochure | Speaker Biographies
THURSDAY, JANUARY 22
11:30 am Conference Registration
12:30 pm Luncheon Presentation: Looking to Achieve Amazing TPP Yields in CHO and HEK293? Introducing FectoPRO™, a Novel Powerful Transfection Solution
Mathieu Porte, MSc, Bioproduction Project Leader, R&D, Polyplus-transfection®
Low transfection efficiency of CHO cells is a major bottleneck hampering Transient Protein Production (TPP). Polyplus-transfection®, with its 10+ year expertise in transfection, has developed a novel technologically advanced transfection solution specifically designed for bioproduction. FectoPRO™ outperforms currently available PEI-based and lipid-based transfection reagents. We will present data and protocols leading to unmatched protein and antibody yields in CHO and HEK-293 cells.
1:30 Ice Cream Break in the Exhibit Hall with Poster Viewing
2:00 Chairperson’s Opening Remarks
Dominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc.
2:05 Application of UPR Response and Cell Cycle Checkpoint Engineering for CHO Cell Line Construction
Takeshi Omasa, Ph.D., Professor, Institute of Technology and Science, University of Tokushima
The construction of high-producing CHO cells is an important issue for therapeutic protein production. We introduce our new approach for Unfolded Protein Response (UPR)-based cell line construction and accelerating transgene amplification system using cell cycle checkpoint engineering.
2:45 Protein Production by PEI-Mediated CHO Transfection
Yves Durocher, Ph.D., Team Leader, Protein Production, Biologics and Subsequent Entry Biologics, Life Sciences - NRC Human Health Therapeutics Portfolio, National Research Council Canada
We present data describing our CHO-EBNA1 transient expression platform for the production of various proteins. For more difficult-to-express proteins or proteins needed in large quantities, we also developed an inducible CHO pool platform that allows the generation of stable pools expressing high levels of monoclonal antibodies in less than 3 weeks post-transfection. This platform can also be used to derive stable CHO clones for manufacturing therapeutic r-proteins candidates.
3:15 Systems-Based Approaches for Enhancing Transient Protein Expression in CHO Cells
Jonathan Zmuda, Ph.D., Associate Director, R&D, Thermo Fisher Scientific
CHO cells are known to express lower levels of transient proteins than HEK293, but are desirable for biopharmaceutical development since they are the most commonly used cell type for bioproduction. To address the need for a higher-expressing transient CHO platform, systems-based approaches were utilized whereby the latest advances in cell culture media and feeds, transfection reagents and enhancers were optimized through multifactorial DoE to generate a complete solution for enhanced protein expression with titers comparable to HEK293-based systems.
3:45 Rapid Protein Production with Flow Electroporation: Higher Titers, Cell Type Flexibility and Scalability
James Brady, Ph.D., MBA, Director, Technical Applications, MaxCyte, Inc.
As protein therapeutics grow, future success will depend on the ability to be flexible regarding cell type and the ability to quickly scale up or scale down. In this presentation, data from MaxCyte flow electroporation will be presented demonstrating its scalability, high transfection efficiency and cell viability of commonly used cells, including CHO-S, HEK293 and insect cells, and production of antibody titers >1 g/L in 2 weeks.
4:15 Refreshment Break in the Exhibit Hall with Poster Viewing
5:00 High-Throughput mAb Expression and Protein A Purification Platform Based on Transient CHO
Yashas Rajendra, Ph.D., Research Scientist, Biotechnology Discovery Research, Eli Lilly and Company
We developed a PEI-mediated transient CHO-GS KO expression system that can generate mAb titers up to 0.5 g/L. We also developed a semi-automated Protein A purification process. Using a single liquid handling robot, up to 72 unique mAbs can be simultaneously purified from 24DWP (deep-well plate) transfections. This process yields 0.3 mg-1 mg of high-quality mAb at concentrations >0.5 mg/ml in a standard formulation buffer, enabling rapid characterization of mABs in multiple assay formats.
5:30 PANEL DISCUSSION: CHO Cells: Transient, Stable or Both
Recombinant protein expression in mammalian producer cells such as CHO offers significant advantages in protein quality and post-translational modification. Expression can be transient, which achieves fast process times and high yields, or stable, which takes greater time to convert but offers other gains. This panel convenes experts to share insights into the benefits and tradeoffs of meeting the demand for recombinant proteins by engineering transient or stable cell lines or a combination of both.
Moderator: Dominic Esposito, Ph.D., Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc.
James Brady, Ph.D., MBA, MaxCyte, Inc.
Yves Durocher, Ph.D., National Research Council Canada
Takeshi Omasa, Ph.D., University of Tokushima
Yashas Rajendra, Ph.D., Eli Lilly and Company
Jonathan Zmuda, Ph.D., Thermo Fisher Scientific
6:00-7:00 Reception at the Tiki Pavilion
Day 1 | Day 2 | Download Brochure | Speaker Biographies