PepTalk 2017
PepTalk 2017
Archived Content

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Fine-Tuning Expression in CHO
Turning the Work Horse Into a Race Horse
January 23-24 


Day 1 | Day 2Download Pipeline 4 Brochure 

CHO can be finicky and difficult to manage if the strain choices, culture conditions, etc. are not carefully controlled. This conference will present methods to not only utilize CHO cells more effectively, but also to turn the work horse into a race horse by improving expression times and yields.


Buzz Session Info  

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1:30 pm Conference Registration

2:00 BuzZ Session A

2:45 Refreshment Break in the Exhibit Hall with Poster Awards

3:30 BuzZ Session B

4:15 End of Day

4:15-4:30 Registration for Dinner Short Courses

4:30 – 7:30 Concurrent Dinner Short Courses (SC5-9)*

*Separate Registration Required


7:30 am Conference Registration and Morning Coffee

Harnessing CHO for Success 

8:15 Chairperson's Opening Remarks

Susan T. Sharfstein, Ph.D., Associate Professor, College of Nanoscale Science and Engineering, University at Albany - State University of New York 



8:20 High-Level Protein Production in CHO Cell Pools Using the Cumate Gene-Switch with Lentiviral Vectors or with an Optimized Transfection Protocol

Bernard MassieBernard Massie, Ph.D., Director, R & D, Human Health and Therapeutics Portfolio, National Research Council Canada

Fast and efficient protein production for structural and functional studies is a crucial issue for research and industry. We will present here that using the powerful cumate-regulated promoter which, in addition to be inducible, is the strongest promoter we have tested thus far in CHO cells, pools of CHO cells stably expressing high-level of recombinant proteins (>200 mg/L) can readily be generated in less than 6 weeks with either lentiviral vectors or following an optimized transfection protocol.

9:00 Metabolic Engineering of CHO Cells for Production of Heparin

Susan T. Sharfstein, Ph.D., Associate Professor, College of Nanoscale Science and Engineering, University at Albany - State University of New York

We hypothesized that HP could be produced in CHO cells by metabolic engineering and have engineered in two critical enzymes, NDST-2 and 3OST-1. The resulting cells show increased anticoagulation activity, but do not produce pharmaceutical heparin. Current efforts are focused on fine-tuning enzyme activities and balancing endogenous and exogenous genes to improve the GAG structures. Results from these studies may also apply to engineering of glycan structures on therapeutic glycoproteins.

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

Woo Suk Ahn, Ph.D., Post Doctoral Associate, Department of Chemical Engineering, Massachusetts Institute of Technology

Metabolism of CHO cells changes significantly in culture as cells transition from growth phase to stationary phase. In this talk, I will highlight progress on 13C-metabolic flux analysis (MFA) and dynamic-MFA. Application of these new tools may allow identification of intracellular bottlenecks at specific stages in CHO cell cultures and eventually lead to novel strategies for improving metabolism and optimizing biopharmaceutical process performance.

10:00 Coffee Break in the Exhibit Hall with Poster Viewing

10:45 MicroRNAs and Apoptosis in CHO Cell Culture

Joseph Shiloach, Ph.D., Chief, Biotechnology Core Laboratory, NIDDK, NIH

This study determined changes in microRNA (miRs) expression in Chinese hamster ovary (CHO) cells undergoing apoptosis induced by exposing the cells to nutrient-depleted media. Inhibition of the mmu-miR-466h increased the expression levels of those genes and resulted in increased cell viability and decreased Caspase-3/7 activation. The up-regulation of mmu-miR-466h in response to nutrient depletion causes inhibition of several anti-apoptotic genes in unison.

11:15 Model-Based Optimization of CHO Cell Culture with Intermittent Harvesting

Saeideh Naderi, Ph.D., Researcher, Department of Chemical Engineering, University of Waterloo

A comprehensive dynamic model was developed and used for an offline model-based optimization of MAb production with the goal of enhancing productivity through intermittent refreshment of the culture media. Experimental implementation of this model-based strategy produced a MAb yield in a 3-step intermittent harvesting operation which was approximately twice the productivity obtained in a batch operation of the same overall duration.

11:45 Genetic and Metabolic Profiling of Monoclonal Antibody:  Production Cell Lines

Sohye Kang, Ph.D., Principal Scientist, Product Attribute Sciences, Amgen Inc. 


 12:15 pm Close of Morning Session 

12:30 Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own


Improving Yields and Engineering Overexpression  

2:00 Chairperson's Remarks

Kelvin H. Lee, Ph.D., Professor, Chemical and Biomolecular Engineering, University of Delaware  

2:05 Russell Body-Inducing Propensity of Individual IgG Clones and Overexpression Fitness

Haruki Hasegawa, Ph.D., Senior Scientist, Protein Science, Amgen, Inc.

Russell bodies (RBs) are intracellular aggregates of immunoglobulins frequently found in the plasma cells of B lymphoproliferative disorder patients. Using recombinant expression systems, we show that individual IgG clones possess distinctive RB inducing propensities that can surface differently under normal and abnormal cellular conditions. We explore the potential value of imaging-based RB detection assays to identify IgG clones with low over-expression fitness.

2:35 Co-Overexpression of Alanine Aminotransferase 1 (ALT1) in Chinese Hamster Ovary Cells Overexpressing Taurine Transporter (TAUT) Further Stimulates Metabolism and Enhances Product Yield

Hisahiro Tabuchi, Ph.D., Chief Scientist, API Process Development (Biotechnology), Pharmaceutical Technology, Chugai Pharmaceutical Co., Ltd.

Our cell engineering strategy has unique potential for the improvement of mAb-producing cells. TAUT-overexpressing cell lines rapidly accumulated the by-product alanine, and our challenge was to apply a strategy for modulating cell metabolism to utilize this by-product to achieve a high mAb yield. To accomplish this, we genetically modified the DXB11/TAUT/mAb1 strain to co-overexpress ALT1. The co-overexpressing strain gave a higher mAb yield.

MaxCyte3:05 Fully Scalable Transient Gene Expression in CHO Cells for Gram-Scale Antibody Production

James Brady, Ph.D., Director, Technical Applications, MaxCyte

Transient gene expression (TGE) directly within CHO cells can greatly shorten the timeline of antibody development by eliminating the need for transient HEK systems. Data is presented within this talk demonstrating rapid gram-scale antibody production using the MaxCyte® STX™ Scalable Transfection System. Secreted antibody titers exceeding 400 mg/L allow for production of over a gram of antibody from a total culture volume of only two liters following a single CHO cell transfection.

3:35 Sponsored Presentation (Opportunity Available)

3:50 Refreshment Break

4:15 Proteomic Analysis Identifies Cofilin as a Metabolic Engineering Opportunity for Enhanced Productivity

Kelvin H. Lee, Ph.D., Professor, Chemical and Biomolecular Engineering, University of Delaware

We performed a proteomic analysis of a series of methotrexate amplified CHO cells overexpressing a recombinant protein and observed decreasing cofilin expression with increasing protein secretion. Based on this observation, we developed an RNAi strategy to alter cofilin expression and found that cofilin knockdown results in increased recombinant protein secretion in adherent and suspension cells.

4:45 Growth within High Density Micro-Environments Stimulates Increased Yields of CHO-Produced Biologics

Kevin Sunley, Ph.D., Senior Scientist, XBiotech USA, Inc.

CHO cells localized to micro-environments within commercially available macroporous microcarriers were observed to increase their cell-specific productivity (Qp) proportionally with an increase in the cell density within their immediate local environment. Resulting in a multi-fold increase in product yields over a comparable stirred-tank suspension culture; the role of the localized culture micro-environment, independent of total culture cell density will be discussed.

5:15 Close of Session

5:30-6:30 Networking Reception in the Exhibit Hall with Poster Viewing

6:30 Close of Day

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