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Cambridge Healthtech Institute’s 3rd Annual
Applying Expression Platforms
Transient, Stable or Both?
January 21-22, 2016

Speed, limiting risk and protein quality are often cited as advantages of transient protein production (TPP), while stable transfection – the longer and more complex process – has the advantage of producing long-term expression of the protein of interest. The rapidly increasing need for recombinant proteins necessitates further improvements in both technologies.

Cambridge Healthtech Institute’s 3rd Annual Applying Expression Platforms conference convenes protein expression specialists who share their experiences with the differences, tradeoffs, advantages and improvements in producing recombinant proteins in transient and stable production systems, along with the systems’ advantages and disadvantages and when to use both.

Final Agenda


7:45 am Conference Registration and Morning Coffee


8:55 Chairperson’s Opening Remarks

Christopher W. Kemp, Ph.D., President, Kempbio, Inc.

9:00 A Comparison of Two Methods for the Gram-Scale Production of Recombinant IgG in Transiently Transfected Cultures of HEK-293 and CHO-S Cells

Chris_KempChristopher W. Kemp, Ph.D., President, Kempbio, Inc.

The use of transient transfection to produce significant quantities of recombinant immunoglobulins and other proteins is an important tool for development of new therapeutics and diagnostics. Transfections mediated by PEI and transductions mediated by BacMam viruses are diverse methods for the introduction of genes into cells at scale. This presentation compares the methodology and results for production of rIgG using these methods in expressions conducted using HEK-293 and CHO-S cells.

9:30 Respiratory Syncytial Virus-Like Particles (RSV VLPs) Made by Transient Transfection of HEK293 Cells Protects the Lower as Well as the Upper Respiratory Tract

Pramila_WalpitaPramila Walpita, Ph.D., Assistant Professor, Department of Tropical Medicine, Medical Microbiology and Pharmacology, John A. Burns School of Medicine, University of Hawaii at Manoa

In vitro studies demonstrated that these suspension-adapted transiently transfected RSV VLPs were functionally assembled and immuno-reactive. In vivo studies in cotton rats (CRs) showed that after two doses of these VLPs given three weeks apart induced potent neutralizing antibody response and protected both the lung and the nose when challenged with live RSV A2 virus.

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

11:00 Transient Expression at Genmab: How to Keep the Engine Running

Tom_VinkTom Vink, Ph.D., Associate Director, Cell & Molecular Science, Genmab BV

The ever-increasing demands in volume and speed of our transient expressions has driven us to optimize both our process and our technologies. This presentation describes our recent improvements in this production process, using automation and linear expression elements to generate large panels of antibodies and optimizing each process step to decrease time lines.

11:30 HEK293 & CHO New Screening Tools, Tips and Tricks for Increasing Expression

Sam Ellis, Vice President & Biochemist, Thomson Instrument Company

The conditions for HEK293, CHO need to be scalable at small scale 15mLs- up to medium production at 3L . Data will be presented on techniques and technology that allow for mimicking large scale fermentation with non-controlled devices . All of these techniques will be given with existing case studies for technologies involving protein or antibody production, structural biology, and can lead to successful transfer from different protein groups.

12:00 pm Session Break

Icosagen12:15 Luncheon Presentation: A Stable Episomal Expression System in CHO for Mammalian Protein Production

Kadaja_MeelisMeelis Kadaja, Ph.D., MBA, Director, Business Development, Icosagen Technologies Inc

We have developed a novel technology to stably maintain expression vectors in dividing mammalian cells. This enables efficient and scalable production of recombinant proteins with low endotoxin levels in up to gram quantities. Our system is suitable for the generation of production cell banks in 10 days, and for the manufacturing of recombinant antibodies. In addition, we show that our expression system can be tailored to improve the quality of the recombinant protein.

1:15 Ice Cream Break in the Exhibit Hall with Poster Viewing


2:00 Chairperson’s Remarks

Tharmala Tharmalingam, Ph.D., Scientist, Process and Product Development, Amgen, Inc.

2:05 New Workflow Platform for High-Throughput Cell Line Development

Christian_BenderChristian Bender, Ph.D., Computational Biologist, Antibody Lead Discovery, Bayer HealthCare

We present a new workflow system automating the Cell Line Development process for mammalian cells used to produce therapeutic antibodies and proteins. It tracks clones produced and screened in high-throughput mode, collects relevant characterization data and streamlines high-throughput workflows by interfacing with automation equipment and bioreactors. We show how Bayer’s Global Biologics groups use it and present applications of the generation of GMP-ready cell lines and their transfer into process development.

2:35 Epigenetic Marks as Early Indicators of Production Stability of Recombinant CHO Cells

Ulrich_GoepfertUlrich Goepfert, Ph.D., Principal Scientist, Cell Line & Molecular Development, Roche Innovation Center Penzberg

During expansion and maintenance, CHO cell lines are prone to production instability, which may be caused by promoter silencing, loss of transgene copies or posttranscriptional effects. Silencing of recombinant genes may be accompanied by DNA methylation and histone modification. We examined a variety of epigenetic modifications and identified molecular indicators which provide the opportunity to enrich stable producers.

3:05 Next Level of E. coli Expression - Optimized Expression of Complex Biosimilars

Vikström_DavidDavid Vikström, Ph.D., CTO, Xbrane Bioscience

3:35 Refreshment Break in the Exhibit Hall with Poster Viewing

4:15 Comparison of ExpiCHO and Expi293 Expression Systems: Considerations for Transient Protein Production

Nina_JainNina Jain, Research Associate, Alexion Pharmaceuticals

We compare Thermo Fisher Scientific’s high-density transient transfection systems, Expi293 and ExpiCHO. We discuss the expression and characterization of multiple proteins (mAbs, Fabs, bispecifics and fusion proteins) generated in both systems. We also compare the proteins expressed in these systems to reference standards generated in stable production cell lines and discuss the implications this data has for system selection.

4:45 A High Cell Density Transient Transfection System for Therapeutic Protein Expression Based on a CHO GS-Knockout Cell Line

Gavin_BarnardGavin Barnard, Ph.D., Group Leader, Eli Lilly and Company

We discuss the development of a PEI-mediated transient CHO expression system capable of generating high titers, scalable up to 6L. This was achieved through rigorous optimization of cell density, DNA and PEI concentrations followed by process development strategies. The system was further improved by the co-expression of a specific transcription factor (XBP1S). This platform enables rapid expression of therapeutic protein candidates, thus alleviating one bottleneck in the drug development process.

5:15 Molecule Assessment and CLD for mAb Candidate Selection

Tharmala_TharmalingamTharmala Tharmalingam, Ph.D., Scientist, Process and Product Development, Amgen, Inc.

Molecule Assessment is the consideration of manufacturing, stability and product attributes when designing/selecting protein candidates for commercialization. This is done at the Cell Line Development stage not only to facilitate meeting First in Human needs but also to ensure productivity needs to meet future commercial demands.

5:45 Close of Day

6:00-7:00 Reception in the Tiki Pavilion



8:00 am Conference Registration and Morning Coffee


8:30 Chairperson’s Remarks

Richard Altman, MS, Research Scientist, Discovery Research, Alexion Pharmaceuticals

8:35 Challenges with Cell Line Development for Recombinant Coagulation Factors – Flexibility Is Key

Arna_AndrewsArna Andrews, Ph.D., Director, Cell Line Development, CSL Limited

The manufacture of recombinant antibody therapeutics has evolved dramatically over the last decade largely due to the fact that antibodies are generally well-behaved molecules compatible with a platform process approach. In contrast, these benefits are not generally applicable to non-antibody molecules, such as recombinant coagulation factors where higher degrees of molecule complexity lead to increased protein quality criteria and the need for more flexible approaches to cell line selection and process development.

9:05 Transient Production in Nicotiana and Its Role in the Development of an Anti-Ebola Virus Monoclonal Antibody Cocktail

Larry_ZeitlinLarry Zeitlin, Ph.D., President, Mapp Biopharmaceutical and LeafBio, Inc.

The pros and cons of transient production in Nicotiana will be presented with a focus on the unique development path of a monoclonal antibody cocktail experienced during the 2014-2015 Ebola outbreak in West Africa.


9:35 The Flexibility and Throughput of a Transient Protein Production Core Group Efficiently Supports the Protein Demands for Early Drug Discovery Studies

Richard_AltmanRichard Altman, MS, Research Scientist, Discovery Research, Alexion Pharmaceuticals

A robust, reproducible transient protein production facility provides critical support to drug discovery efforts. We review the genesis and evolution of our scalable transient protein production efforts. The impact to our overall drug discovery efforts will be discussed through case studies highlighting the variety of applications (functional studies, reagent production, etc.) and scale (microtiter plate to WAVE bioreactor) our system provides. We also discuss improvements to our throughput by automation.

10:05 Coffee Break with a Poster Pavilion

PepTalk is proud to support and recognize the protein scientists of tomorrow during the Poster Pavilion. This time has been set aside to view the Student Fellowship posters and interact with presenters one on one. This opportunity gives job seekers the chance to share their expertise with future/potential employers or develop contacts to further their research.


⊲ Featured Presentation
11:00 Enhanced Protein Purification Efficiency through Automation and Laboratory Design

Kenneth_WalkerKenneth Walker, Ph.D., Scientific Director, Biologics, Amgen, Inc.

In order to keep pace with the high-throughput cloning and expression needed to screen large panels of therapeutic candidates, we developed systems to substantially increase protein purification throughput through enhanced laboratory design and the use of novel chromatography instrument automation. With minimal user intervention, these systems can process a wide array of samples employing flexible, advanced chromatography methods that efficiently generate high-quality products.

11:30 Automated High-Throughput Protein Purification Workflows for Discovery of Novel Therapeutics

Avinash_GillAvinash Gill, Ph.D., Scientific Manager, Antibody Engineering, Genentech, Inc., a member of the Roche Group

In combination with automated HTP workflows for antibody expression and purification, an efficient process has been put in place for making large numbers of purified antibody variants available to be screened in biological/functional assays, epitope identification and manufacturability evaluation. Expression and purification are carried out at 2 scales of cell culture – 1mL (deepwell blocks) and 30 mL (tubespin columns), to generate antibodies, antibody fragments and other therapeutic protein formats.

Friday, January 22, 12:00 pm

Protein therapeutics is one of the fastest-growing global markets, driven by increasing adoption of protein- over non-protein drugs, growing funding for protein engineering and reduced drug discovery timelines and costs. As the science improves, so does the complexity of the R&D organization: it really does “take a village” to bring nextgeneration therapies to market and patients who need them. Ensuring product quality plus speed to market requires collective insights from experts working across the stages of protein science R&D – as embodied by panelists representing each PepTalk Pipeline topic.

They discuss:

  • Highlights from the week’s Pipeline presentations
  • What’s next for protein therapeutics?
  • How to prepare for and solve these challenges


Danny_ChouDanny Chou, Ph.D.

former Senior Research Scientist, Biologics Development,
Gilead Sciences; President and Founder, Compassion BioSolution


Dominic EspositoDominic Esposito, Ph.D.

Director, Protein Expression Laboratory,
Frederick National Laboratory for Cancer
Research, Leidos Biomedical Research, Inc.

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Randall BrezskiRandall Brezski, Ph.D.

Scientist, Antibody Engineering, Genentech, Inc.

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Marisa JoubertMarisa K. Joubert, Ph.D.

Senior Scientist, Process & Product
Development, Amgen, Inc.

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Rakesh DixitRakesh Dixit, Ph.D.

DABT, Vice President, Research &
Development; Global Head,
Biologics Safety Assessment, Medimmune

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Jonas SchaeferJonas V. Schaefer, Ph.D.

Head, High-Throughput Laboratory,
University of Zurich

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Thomas LaueThomas Laue, Ph.D.

Professor, Biochemistry and Molecular
Biology; Director,
Biomolecular Interaction Technologies
Center (BITC), University of New Hampshire

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1:15 Close of Conference