Archived Content

EXPRESSION STREAM  

2nd Annual  

Optimizing Protein Expression May 2 - 3


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WEDNESDAY, MAY 2

7:00 am Registration and Morning Coffee

 

PROMOTING PROTEIN PRODUCTION 

8:30 Chairperson’s Opening Remarks

Krista Bowman, Ph.D., Senior Scientific Manager, Structural Biology, Genentech, Inc.


» 8:40 Keynote Presentation 

Approaches to Maximize Protein Expression for Discovery and Development Efforts

William H. BrondykWilliam H. Brondyk, Ph.D., Senior Scientific Director, Therapeutic Protein Discovery, Genzyme – A Sanofi Company (biography)

Achieving sufficient expression levels of recombinant proteins to meet the needs for both discovery and development efforts are a goal for biotechnology and pharmaceutical companies. In this talk, a description of our currently used insect and mammalian expression systems will be presented. Specific information will be discussed regarding our optimization of vectors, strategy for the selection of the expression system, methods for generating high expressing CHO pools and clones, and gene design.


» 9:10 FEATURED PRESENTATION 

Transient Expression of Proteins in HEK293 and CHO Cells –
or is There an Alternative?

Sabine GeisseSabine Geisse, Ph.D., Director, NLS, Novartis Pharma AG (biography)

We describe a novel cell line, CAP-T® derived from human amniocytes, as host in transient protein production, and the establishment of suitable protocols for transfection and expression of recombinant proteins. Moreover, we show comparative analyses of expression to the well-known HEK293 and CHO transient protein production platforms. In brief, the addition of the CAP-T® cells to the repertoire of cell lines amenable to transient protein production clearly enhances the chances of success.


9:40 Transient (HEK293) and Stable (CHO) Expression of Dual Variable Domain (DVD) - Ig™ Molecules

Gerald CarsonGerald Carson, Senior Principal Scientist, Biologics, Abbott Bioresearch Center, Inc. (biography)

  • Factors influencing drug-like properties (DLP) of DVD-Igs 
  • Transient (HEK293 cells) and Stable (CHO cells) expression of DVD-Igs 
  • Modulating inner target binding domain function 

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

 

PROTEIN EXPRESSION IN CHO CELLS 

11:10 An Update on the International Community’s Efforts at CHOgenome.org

Kelvin LeeKelvin H. Lee, Gore Professor of Chemical Engineering & Director, Delaware Biotechnology Institute, University of Delaware (biography

Today, a quarter of all FDA approved new drugs are biopharmaceuticals, most of which are produced in Chinese hamster ovary (CHO) cells. The CHO K1 cell line is an ancestor to many production cell lines and was recently fully sequenced. We will discuss the aspects of the international community’s efforts at developing an infrastructure to support, host, and disseminate genome-scale data related to CHO cell lines.

11:40 Dynamic Model for CHO Cell Engineering     

Kyongbum LeeKyongbum Lee, Ph.D., Associate Professor & Acting Chair, Chemical and Biological Engineering, Tufts University (biography)

CHO cell fed-batch processes have progressed significantly over the past decade, with protein titer consistently reaching the gram per liter level. Progress has largely resulted from separate advances in process and cell line development. We use a dynamic model to explore~104 combinations of process and cell modifications. Knockdowns in glycolysis were the most beneficial; however, depending on the process conditions, such knockdowns could reduce the antibody titer. Our results highlight the need to consider process and cell modifications together.

Catalent12:10 pm Luncheon Presentation I
Protein Expression in CHO Cells: Comparison of GPEx® to a Traditional Cell Line Engineering Technology
Gregory Bleck Gregory T. Bleck, Ph.D., Research and Development, Platform Lead-Biologics, Catalent Pharma Solutions (biography)Speed, cell line stability, high productivity and excellent protein quality are the goals of any mammalian cell line engineering technology, but does the way cell lines are made have any impact on their ability to produce a hard to express protein?  GPEx® is a versatile protein expression system which is capable of transferring genes of interest into a wide variety of mammalian host cells.  A comparison was performed between GPEx® and a traditional cell line engineering technology.  An antibody that was easy to express using traditional methods and three molecules that were poor producers and/or had poor cell line stability using traditional methods were analyzed.  Cell lines expressing the four products were developed using both traditional methods and GPEx®.  No significant difference was observed for the easy to express antibody between the two approaches, however for the other three problem molecules, GPEx® equivalent cell lines outperformed in both protein production and stability.  Results of the analysis will be discussed
 

12:40 Luncheon Presentation II (Sponsorship Opportunity Available) or Lunch on Your Own

 

PROTEIN EXPRESSION IN CHO CELLS AND AN ALTERNATIVE SYSTEM 

1:30 Chairperson’s Remarks

Anne London, Ph.D., Investigator II & Lab Head, Mid-Scale Protein Production, Novartis Institutes for BioMedical Research, Inc.

1:35 Maximizing Production of Soluble IL-15: Cytokine Receptor Superagonist Complexes by CHO Cells

Peter Rhode, Ph.D., Vice President, R&D, Altor BioScience Corp. (biography)

Interleukin-15 is a promising immunostimulatory cytokine for treatment of cancer and viral diseases. However, its development has been limited by poor expression in bacterial and mammalian systems. We have found that co-expression of an IL-15 superagonist variant with a soluble IL-15 receptor alpha - IgG1 Fc fusion leads to high level production of a fully active IL-15:IL-15Rα complex by CHO cells. A simple, scalable affinity and ion exchange chromatography method was conducted to highly purify this complex, enabling its clinical development.

2:05 The Microalgal Cell Line PTA: A Novel Host for the Expression of Therapeutic Glycosylated Proteins

Alexandre Lejeune, Ph.D., Business & Innovation Director, Algenics (biography)

The increasing importance of biologics as well as knowledge gained from decades of therapies led to challenges and opportunities in the field of biomanufacturing. The technological platform AlgebiosysTM developed by Algenics leverages microalgae capabilities to achieve consistent and qualitative glycoproteins expression. To demonstrate the versatility offered by our microalgal cell line, proofs of concept including monoclonal antibodies and recombinant viral antigens will be presented with emphasis on glycosylation properties.

Polyplus small logo2:35 Transient Gene Expression: So You’re Already Using PEI (polyethylenimine) – What Could be Better?

Habib Horry, Ph.D., Strategic Marketing Manager, Polypus-transfection 

With 10 years of “PEI for transfection” manufacturing experience, Polyplus-transfection® introduces the last generation of PEI for cost effective production of milligrams to grams amount of r-proteins in suspension-adapted mammalian cell lines.

PerkinElmer NEW 20092:50 High Throughput Analysis of Protein Quality in Biotherapeutics Development using Microfluidic-Based TechnologyBrian Gerwe, Ph.D., PerkinElmer, Inc. 

3:05 Refreshment Break in the Exhibit Hall with Poster Viewing 


3:50 Problem Solving Breakout Discussions

Concurrent problem solving breakout discussions, open to all attendees, speakers, sponsors, and exhibitors, provide a forum for discussing key issues and meeting potential collaborators. Plan to take part and explore these topics in-depth. Please pick a topic of your choice, find your table and join in.

7.  Transient Protein Expression – Where Do We Go In The Future?

Moderator:  Sabine Geisse, Ph.D., Director, NLS, Novartis Pharma AG

• Advantages and drawbacks/bottlenecks of current processes
• Ways to improve expression rates and processes
• Can transient expression replace GMP-conform manufacturing cell lines in the future?
• What would be needed to develop and implement above mentioned application of transient production?

8.  Discussion of Real-Life Experiences with Gene Construct Modifications, Cell Line Exchanges or Alterations, or Changes in Cell Line Development Process that have Yielded Improved Protein Production in Mammalian Cell Lines 

Moderator:  Gregory T. Bleck, Senior Director, Research and Development, Catalent Biologics, Catalent Pharma Solutions

• Gene construct modifications (ex. Codon optimization, translation enhancers, new promoters, etc)
• Cell line exchanges or alterations (ex. CHO, 293 HEK, NS0, addition of processing enzymes, etc.)
• Changes to cell line development process (ex. DHFR amplification, GS amplification, single copy gene insertion, viral gene insertion, genetic insulator systems, etc.)

9.  Experiences with Current Recombinant Protein Production Methodologies

Moderator:  Ciarán Cronin, Ph.D., Head, Parallel Protein Production Group, Pfizer, Inc., San Diego

• Vector construction – outsourcing, codon optimization
• Expression hosts, particularly E. coli, insect and mammalian cells
• Baculovirus-infected insect cell expression – current approaches
• Purification tags, tag cleavage enzymes and purification methods

10.  Refolding Proteins: An Answer or Just Another Problem?

Moderator:  Robert M. Petrovich, Ph.D., Head, Protein Expression Core Facility, Laboratory of Structural Biology, National Institute of Environmental Health Sciences, NIH

• Which methods work (rapid dilution, vs dialysis, vs high pressure) best?
• Can you easily and cheaply scale the process?
• Can you reliably refold without an activity assay?
 

11.  How do Genetic Insights (i.e., CHO genome) Help with Protein Expression? 

Moderator:  TBD 

12.  What do ‘Alternative’ Systems Offer, and How do They Perform? Exploring Yeast, Transgenics & Chimerics 

Moderator:  TBD 

13.  Tricks of the Trade: How to Transform/Maintain a Multi Expression System Facility 

Moderator:  TBD 

14.  The Future of Protein Expression in CHO cells 

Moderator:  TBD 

15.  HTP, Robotics & Automation: Cost, Implementation vs. Results 

Moderators:  Brian Gerwe, Ph.D., Account Manager, Caliper, a PerkinElmer Co.
Jeremy Lambert, Manager, Automation Applications & Genomics Marketing, Caliper, a PerkinElmer Co.
 

 

4:50-6:00 Networking Reception in the Exhibit Hall with Poster Viewing

Emerald BioStructures Poster Awards Sponsored by

 

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