PepTalk 2017
PepTalk 2017
2014 Archived Content

Cambridge Healthtech Institute’s Inaugural
Transient Protein Production
From Small-Scale to Large-Scale
January 16-17, 2014

The advantages of transient protein production are recognized. 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) offers significant advantages in protein quality and post-translational modification. However, the rapidly increasing need for recombinant proteins necessitates further improvements in transient protein production technologies.

Cambridge Healthtech Institute’s Inaugural Transient Protein Production meeting convenes protein expression specialists to share their experiences of the differences, tradeoffs, advantages and improvements in producing recombinant proteins in transient production systems.

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


1:00-1:45 pm Conference Registration

Cell Lines: New Developments 

2:00 Chairperson’s Opening Remarks

Athena Wong, Ph.D., Scientist, Early Stage Cell Culture, Genentech, Inc.

Keynote Presentation

2:05 Recombinant Protein Production in Transient Fashion: A Mature Technology?

Sabine GeisseSabine Geisse, Ph.D., Director/NLS, Novartis Institutes for BioMedical Research

The huge wealth of recombinant proteins generated by transient expression, as well as the availability of well-documented protocols in the public domain, reflects the remarkable success of transient protein production over the past 15 years. Thus, the term “mature technology” seems justified – yet, process gaps and shortcomings do exist. The presentation will cover a brief overview on our currently applied routes to protein production (including the novel CAP-T™ expression system) and address some of the issues encountered and future perspectives.

2:45 Application of a New Human Cell Line, F2N78, as a Host Cell for Transient Production of Biopharmaceuticals

Jong-Mook KimJong-Mook Kim, Ph.D., Director, Cell Science Team, R&D Division, Celltrion Inc.

F2N78, generated by a somatic fusion of 293 cells with Namalwa cells, showed a constitutive expression of EBNA1, and human-specific glycosylation enzymes, GnTIII and α(2,6)ST, for over one year under serum-free suspension culture conditions. Monoclonal antibody (mAb) productivity of ~100 μg/ml was obtainable six days post-transfection with oriP expression vector and the produced mAb could be applied for a selection of proper mAb during early development process of a new target molecule.

3:15 Use of an Anti-Apoptotic Cell Line for High-Throughput Transient Gene Expression

Athena WongAthena Wong, Ph.D., Scientist, Early Stage Cell Culture, Genentech, Inc.

This presentation describes the development of a polyethylenimine (PEI) transient transfection system using an anti-apoptotic CHO-K1 host cell line generated by deleting two pro-apoptotic factors, Bax and Bak, using zinc finger nuclease-mediated gene disruption. The Bax Bak double knockout (DKO) cells expressed 3-4 fold higher antibody titers than CHO-K1 cells, maintained higher viability post-transfection and had decreased active caspase-3. The optimized high-throughput process can be used for scales ranging from automated tube spins and shake flasks to stirred tank bioreactors.

MaxCyte3:45 The Impact of Scalable Transient Gene Expression: Maximizing CHO Antibody Production to Accelerate Project Timelines

Brady_JamesJames Brady, Ph.D., Director, Technical Applications, MaxCyte, Inc. 

CHO transient gene expression (TGE) greatly accelerates antibody development by eliminating the need to change cell backgrounds during scale up to biomanufacturing. Data using MaxCyte transient transfection will be presented demonstrating its scalability, the production of antibody titers >1g/L, and the rapid generation of high yield stable CHO cell lines. 

4:00 Refreshment Break in the Exhibit Hall With Poster Viewing

4:45 Rapid Screening of Membrane Protein Expression in Transiently Transfected Insect Cells

Hao ChenHao Chen, Ph.D., Senior Scientist, Protein Technologies, Amgen, Inc.

Membrane proteins play critical roles in many biological processes and are the focus of intense biomedical research. One bottleneck for studying membrane proteins is the difficulty in expressing correctly folded and stable proteins, which often requires time and resource-intensive protein engineering and optimization. Here, we present an ultrasensitive method for rapidly screening membrane protein expression in insect cells, in which only nanogram levels of unpurified proteins are required.

5:15 Optimized Signal Peptides for the Development of High-Expressing CHO Cell Lines

Lars KoberLars Kober, Ph.D., Scientist, Cellca GmbH

Powerful signal peptides: Mammalian cell lines are the first choice for the production of biotherapeutic secretory proteins. In our last article we identified two very potent human-derived signal peptides suitable for the production of biopharmaceutical proteins. With these signal peptides we were able to generate highly productive mammalian cell lines with cell-specific productivities up to 90 pg/cell and day. Furthermore, we have obtained product concentrations up to 4 g per liter in a protein-free chemical defined cell culture medium. 

5:45 Close of Day

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