PepTalk 2017
PepTalk 2017
2014 Archived Content

Cambridge Healthtech Institute’s Sixteenth Annual
Recombinant Protein Expression and Production
Achieving Quality and Quantity
January 15-16, 2014

Biopharmaceuticals currently represent the fastest-growing sector of the pharmaceutical industry driven by a rapid expansion in the manufacture of recombinant protein-based drugs. To meet the demand, it is crucial to increase the throughput of expression, production and purification processes and systems.

Cambridge Healthtech Institute’s Sixteenth Annual Recombinant Protein Expression and Production meeting explores the newest data and innovations relating to the best choices in hosts/systems, as well as ways to “rescue” existing systems and make them work more effectively to produce the quality and quantity of the desired biotherapeutic.

Day 1 | Day 2 | Download Production Brochure | Speaker Biographies 



1:30-2:00 pm Conference Registration

BUZZ Sessions png

2:00 BuzZ Session A (More Details >>)

3:00 Refreshment Break in the Exhibit Hall with Poster Awards

3:45 BuzZ Session B (More Details >>)

4:30-5:00 Short Course Registration

5:00-8:00 Dinner Short Courses (SC8-SC14) More Details >> 




7:30 am Conference Registration

8:00 Morning Coffee

Expression: Improving Yields 

8:15 Chairperson’s Opening Remarks

Karl E. Griswold, Ph.D., Associate Professor, Thayer School of Engineering, Dartmouth College

Keynote Presentation

8:20 Protein Expression Technologies: Evolution or Revolution?

Lorenz M. MayrLorenz M. Mayr, Ph.D., Vice President, Reagents & Assay Development, AstraZeneca, Inc.

Protein expression has long been viewed as a mature discipline, but current trends in drug discovery show an increased demand for fast, efficient expression systems. We will discuss comprehensive and contemporary activities in place for protein expression at AstraZeneca. Topics include an overview of established protein expression technologies; novel technologies and trends for protein expression in drug discovery research; case studies for difficult-to-express proteins and protein complexes; finding the balance between in-house efforts and outsourcing; and a summary and outlook.

9:00 Novel Strategy for Production of Difficult-to-Express Proteins in Escherichia coli Based on an Anchored Periplasmic Expression System

Ki Jun JeongKi Jun Jeong, Ph.D., Assistant Professor, Chemical and Biomolecular Engineering, KAIST

For the efficient production of difficult-to-express proteins such as aggregation-prone proteins and lytic enzymes in the E. coli host, we have developed a new production system based on an Anchored Periplasmic Expression (APEx) system. In this APEx, protein aggregation and lytic activity can be prevented through anchoring of individual proteins to the inner membrane. This concept was successfully demonstrated with two model proteins (aggregation-prone human leptin and lytic lipase).

9:30 Production of Disulfide-Rich Peptides via Expression in the Periplasm of Escherichia coli 

Glenn F. KingGlenn F. King, Ph.D., Research Scientist, Institute for Molecular Bioscience, University of Queensland

Disulfide-rich venom peptides (DRPs) have received attention as potential therapeutics because they potently target a wide range of receptors and their rigid disulfide framework makes them highly stable in vivo. However, expression of functional DRPs in the reducing environment of the E. coli cytoplasm is difficult. Thus, we developed a protocol for DRP expression in the E. coli periplasm, which houses the machinery for disulfide-bond formation. We demonstrate this method for production of a wide range of DRPs (2–8 kDa, 2–6 disulfide bonds).

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

10:45 A Simple PiggyBac Transposon-Based Mammalian Cell Expression System for Inducible Protein Production

James M. RiniJames M. Rini, Ph.D., Professor, Molecular Genetics and Biochemistry, University of Toronto

We have developed a doxycycline inducible PiggyBac (PB) transposase-based mammalian cell expression system which greatly simplifies the generation of stably transfected bulk cell cultures for protein production. It works with both adherent and suspension-adapted cell lines and owing to the efficiency of PB-mediated integration, the system allows for the generation of bulk cell cultures in 96-well format. Our interests are focused on secreted and membrane proteins and the utility of the system will be demonstrated with a number of examples.

Cell Line Development 

11:15 High-Throughput Imaging to Increase the Assurance of Clonality during Cell Line Development

David Shaw, Ph.D., Scientist, Early Stage Cell Culture, Genentech, Inc.

Current methodologies to create monoclonal cell lines include limiting dilution or single-cell sorting at conditions that offer statistical assurance of monoclonality. We are evaluating a fluorescent high-throughput automated imaging workflow that can provide direct evidence on whether the cell line originated from one cell during the cloning step. We will discuss some of the challenges during the development of this protocol and provide case study data.

11:45 Cell Line Development: Can Modifications at the Early Stages Improve Therapeutic Cell Line Selection?

Bernie Sweeney, Ph.D., Senior Group Leader, Mammalian Expression, UCB

The generation of manufacturing cell lines can require screening 100s-1000s of clones. High-throughput technologies have enabled more efficient screening but modifications in early stages of cell line manufacturing may help further. Approaches include the use of engineered cell lines, chromatin modifying elements and overexpression of “helper” proteins. We will present data on some approaches and demonstrate how they can shorten timelines and reduce costs during cell line development.

Selexis small logo12:15 pm Selexis SURE CHO-Mplus™ Libraries: Custom Solutions for Protein Expression Bottlenecks

Pierre-Alain Girod, Ph.D., CSO, Selexis

Ideal protein expression systems should both boost transcription as well as address expression bottlenecks. For the newest SUREtechnology innovation, Selexis leveraged data from the completed SURE CHO-M Genome and Transcriptome project to engineer the SURE CHO-Mplus Libraries designed to address expression issues in CHO-M cells. Selexis’ expression technology doesn’t need antibiotic selection, allowing for screening of multiple auxillary proteins simultaneously. These libraries boost expression over a broad range of difficult-to-express proteins.

Protein Simple12:45 Luncheon Presentation: Why Are You Still Doing Westerns?

Proctor_JohnJohn Proctor, Ph.D., Director, Corporate Development, ProteinSimple

If you are, you must not know about the Simple Western, a gel-free, blot-free, hands-free reinvention of the traditional Western blot. It is a fully automated family of instruments that delivers reproducibility and true quantitation while drastically reducing the hands-on time required when performing a traditional Western. In this presentation, I will illustrate examples of how the Simple Western has been applied in protein expression and production, cell signaling analysis, biotherapeutic characterization and vaccine research.

Purification: Improving Product Quality 

1:50 Chairperson’s Remarks

James M. Rini, Ph.D., Professor, Molecular Genetics and Biochemistry, University of Toronto

1:55 Microbial Production of Folded and Fully Functional Antibacterial Enzymes

Karl E. GriswoldKarl E. Griswold, Ph.D., Associate Professor, Thayer School of Engineering, Dartmouth College

Widespread drug resistance among bacterial pathogens has focused increasing attention on antibacterial biocatalysts and their clinical application. In particular, genetically engineered versions of natural enzymes have the potential to exert powerful therapeutic effects, but scalable production systems, able to service early-stage discovery and development through pilot or production-scale translation, are often lacking. This talk will highlight recent advances that have enabled highly efficient expression and purification of two bactericidal enzymes, one in Escherichia coli and the other in Pichia pastoris.

2:20 A Yeast-Based Platform for High-Yield, High-Quality Production and Purification of Eukaryotic Membrane Proteins

Per Amstrup PedersenPer Amstrup Pedersen, Ph.D., Professor, Biology, University of Copenhagen

Membrane proteins are by far the single group of proteins most difficult to express and purify in a functional form. Engineering expression and solubilization conditions have allowed us to produce high-quality eukaryotic membrane proteins in quantities allowing fundamental protein chemical analysis as well as exploiting their use in industrial applications.

2:45 Optimizing Expression Tags and Fermentation Conditions for Production of Functional Human Integral Membrane G Protein-Coupled Receptor in E. coli 

Alexei YeliseevAlexei Yeliseev, Ph.D., Staff Scientist, Protein Biochemistry, LMBB, National Institutes of Health

Human cannabinoid receptor CB2, a G protein-coupled receptor involved in regulation of immune response, is an important target for pharmaceutical drug development. We optimized production of the functional CB2 receptor, labeling with stable isotopes by fermentation in E. coli, and efficient purification of the receptor by testing expression hosts, cultivation conditions and different combination of affinity tags including C-terminal nanopeptide (Rho-tag). The purified, stable receptor reconstituted into liposomes is amenable to studies by solid-state nuclear magnetic resonance spectroscopy.

3:10 Reversible Labeling of Native and Fusion-Proteins

Michael D. BurkartMichael D. Burkart, Ph.D., Professor, Chemistry and Biochemistry, University of California, San Diego

The reversible covalent attachment of chemical probes to proteins has long been sought as a means to visualize and manipulate proteins. We have developed the full reversibility of post-translational custom pantetheine modification of carrier protein domains and fusion protein for visualization and functional studies. This iterative enzymatic methodology can be used in vitro to reversibly label protein variants for a variety of applications, including isolation, visualization and immobilization.

Pfenex Inc.3:35 Discovery Protein Expression Enabled by Pƒēnex Expression Technology™

Coleman_RussRussell Coleman, Senior Scientist, Pƒēnex Inc.

Discovery scientists’ first step in every product development program is the acquisition of active protein. Leveraging the high-throughput Pƒēnex Expression Technology™ platform, milligram amounts of purified proteins can be produced in support of early stage in vitro and in vivo evaluation. Case studies will provide an overview regarding how Pƒēnex scientists accomplish this task.

3:50 Refreshment Break

4:15 Unfolded Protein Response (UPR) During CHO Cell Production Product Quality

Zhimei DuZhimei Du, Ph.D., Senior Scientist, Cell Sciences & Technology, Amgen, Inc.

We have created a UPR-responsive, fluorescence-based reporter system to detect and quantify specific UPR-mediated transcriptional activation of intracellular signaling pathways in real time without manipulation. The results showed the UPR activation is dynamically regulated during production culture. Also, the clones differed in their UPR induction patterns; the timing and the degree of UPR-induced transcriptional activation were linked to the growth, viability, productivity and product quality of the cells. Additionally, endogenous UPR activation was significantly impacted by the cell culture environment.

4:45 Engineering a Mammalian Cell Line Toolkit that Exhibits Multiple Productivity and Product Quality Profiles

Mark TiéMark Tié, Associate Scientist, Cell Culture Development, Biogen Idec

We have engineered new mammalian host cell lines that broaden the range of achievable product quality attributes. This lets us match a molecule of interest to one of our new host cell lines at the onset of cell line development. We will show data of our in-depth characterizations of these host cell lines using model receptor-FC fusion and monoclonal antibody molecules. Our results demonstrate that different host cell lines can occupy distinct areas of product quality and productivity space.

5:15-6:30 Reception in the Exhibit Hall With Poster Viewing

Day 1 | Day 2 | Download Production Brochure | Speaker Biographies