Cambridge Healthtech Institute’s Fifth Annual

Optimizing Expression Platforms

Tools for Effective Expression, Production & Purification

January 11-12, 2018


The utilization of engineered therapeutic proteins for basic research, clinical diagnostics and therapy continues to expand. Consequently, protein expression laboratory researchers face challenges in efficient expression, production, and purification even while improving quantity and quality and minimizing time and cost. Transient protein production (TPP) has the advantage of speed and limiting risk while stable transfection – the longer and more complex process – has the advantage of producing long-term expression of the biotherapeutic of interest. The rapidly increasing need for recombinant proteins necessitates further improvements in both technologies.

Cambridge Healthtech Institute’s Fifth Annual Optimizing Expression Platforms conference convenes protein expression specialists who share their experiences of the differences, tradeoffs, and improvements in producing recombinant proteins in transient or stable production systems, and who investigate their advantages and disadvantages and when to use both.

Final Agenda


7:45 am Registration and Morning Coffee

Transient Protein Production

8:15 Chairperson’s Opening Remarks

Richard Altman, MS, Scientist, Protein Technologies, Amgen


8:20 Integrating Cell Culture with Magnetic Protein A Bead-Trap Technology Accelerates Antibody Purification

John_KawooyaJohn K. Kawooya, Ph.D., Director, Biologics Optimization, Discovery Research, Amgen

Antibody engineering produces large numbers of molecules (200-500 molecules at 30-50ml each) which require purification, analysis and screening for potency, binding, pharmacodynamics, pharmacokinetics, manufacturability, expression levels and stability in order to select leads. Ever since its inception over 30 years ago, the AKTA system combined with Protein A agarose columns has remained the “workhorse” of antibody purification from cell cultures. However, the inability of this system to process multiple samples in parallel coupled with both its limiting flow rates, its requirement for multiple FTEs to remove cells and particulate from each sample prior to loading together with the potential for sample swapping errors and cross contaminations – all impose major bottlenecks in expediting large purified panels of molecules. In this presentation, I show how a single FTE with parallel Magnetic Protein A bead-trap technology accelerates delivery of high-quality purified antibodies in high yield directly from small (30ml-5liters) to large (25-liter wave-bag) crude cell cultures without centrifugation or filtration.

9:00 Transient Antibody Production: How to Generate Higher Titers

Saurabh_SenSaurabh Sen, Ph.D., Principal Scientist, Biotherapeutics Discovery, Boehringer Ingelheim

The presentation covers topics on optimizing efficient expression and production even while improving quantity and quality and minimizing time and cost. Our Transient Gene Expression (TGE) technology for transient protein production (TPP) has significant advantages by using lesser amounts of coding DNA by 70-80% -- and using 50% less transfection reagent. For research purposes, we have an improved TGE protocol with significant advantage of speed, higher yields and lower costs.

9:30 Advances and Challenges in Transient Plant-Based Therapeutic Protein Production

Karen_McDonaldKaren McDonald, Ph.D., Professor, Chemical Engineering, University of California, Davis

Plants are an excellent host for transient production of therapeutic proteins due to their high expression levels, rapid development timescales, short batch production times, flexibility, robustness, scalability, biosafety, and economics. A variety of transient expression systems/platforms and glycoengineered plant host lines have been developed and commercial-scale facilities have been built. Current work in the area of transient production of recombinant proteins in plant cell suspension cultures will be presented.

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

11:00 Fundamentals of Baculovirus Expression and Applications

Chris_KempChristopher Kemp, Ph.D., President, Kempbio

The baculovirus expression vector system (BEVS) is a major protein expression platform for the production of research and therapeutic grade proteins. BEVS supports the expression of proteins in both insect and mammalian cell hosts and the high efficiency and reproducibility of viral transduction allows the expression of difficult proteins. This presentation focuses on applications of BEVS for the expression of proteins and protein complexes in insect and mammalian cells.

11:30 Accelerated Protein Production via Transient Cell Engineering

Enrique_RotaEnrique Miranda Rota, Ph.D., Research Associate, University College London Cancer Institute.

Biotherapeutic protein development groups are often faced with producing a wide variety of proteins with varying and often complex requirements. Our presentation describes a fully scalable, cGMP compliant, transient cell engineering strategy along with a streamlined purification process that allows us to generate large quantities of purified biotherapeutic proteins quickly to satisfy the needs of our collaborators without having to do labor and time intensive stable clone generation or produce large cell collections.

12:00 pm Session Break

12:15 Luncheon Presentation: New Tools for Screening & Harvesting Solutions for CHO & HEK293 Cells, for Both Transient and Stable Cell

Sam_EllisSam Ellis, Vice President, Thomson Instrument Company

Evaluation of different transfection tools, product quality, and titer for both CHO and HEK293 cell lines. Data will be presented on techniques and technology that mimic large-scale bioreactors in non-controlled devices from 1mL-3L. Technologies presented include well plates and culture tube systems with incorporated filtration methodology. A new direct harvesting technique will also be introduced that eliminates centrifugation while maintaining 0.2um sterile filtration. All of these tools will be presented with case studies from scientists.

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


2:00 Chairperson’s Remarks

Saurabh Sen, Ph.D., Principal Scientist, Biotherapeutics Discovery, Boehringer Ingelheim

2:05 A High-Density CHO-S Transient Transfection System: Comparison of ExpiCHO and Expi293

Tadas_PanavasTadas Panavas, Ph.D., Senior Principal Scientist, Biotherapeutics Molecule Discovery, Boehringer Ingelheim Pharmaceuticals, Inc.

Chinese Hamster Ovary (CHO) cells are the principal mammalian host used for stable cell line generation and biotherapeutic protein production. Until recently, production of milligrams to grams of protein in CHO transient systems was challenging. To overcome such challenges, we evaluated the ExpiCHO system, a high-density CHO-S transient transfection system, and compared it to the Expi293 and FreeStyle MAX CHO transient systems. Detailed analysis was performed on protein titer, monodispersity, enzyme activity, and posttranslational modifications.

2:35 Transient Protein Production: Harmonizing the Process from Construct Generation through Protein Characterization

Richard_AltmanRichard Altman, MS, Scientist, Protein Technologies, Amgen

A robust, flexible transient protein production facility provides critical support to drug discovery efforts. We review the ongoing evolution of our protein production endeavors focusing on two critical components. The first is the strategic assembly of mammalian expression “tools” that gives us a toolbox capable of expressing diverse and challenging candidate proteins. The second is the harmonization of the entire protein production process thereby reducing turnaround times and increasing throughput.


3:05 Difficult to Express Proteins: Novel Plasmid Technology to Significantly Increase Product Yield in CHO Cells

Marco_CacciuttoloMarco Cacciuttolo, Ph.D., Head of Operations, Batavia Biosciences

Yield is still an area that requires significant improvement for many promising recombinant proteins and antibodies. Novel vector technology enables rapid generation of stable, CHO cell lines able to provide at least 10-fold more product per cell.

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

4:15 Lost in Translation: On the Formation of Protein Sequence Variants

Zhongqi_ZhangZhongqi Zhang, Ph.D., Scientific Director, Attribute Sciences, Process Development, Amgen

With modern mass spectrometry and appropriate informatics tools, a large number of low-level sequence variants in therapeutic proteins are detected and quantified. This large collection of information allows for a deeper understanding of the mechanism for the formation of sequence variants, thereby facilitating optimization of cell line and cell culture process to minimize them.

4:45 The Stability of CHO Genome: Essential for Cell Line Characterization or Not?

Noriko_YamanoNoriko Yamano, Ph.D., Senior Scientist, Manufacturing Technology Association of Biologics; Guest Academic Staff, Graduate School of Engineering, Osaka University

The chromosomes in CHO cells frequently cause genomic variations, due to genetic instability. Distribution and stability of chromosomes were examined in CHO-DG44 cells, and two cell lines expressing different numbers of chromosomes were isolated from the original CHO-DG44 cell line to investigate the effect of aneuploid cells on recombinant protein production. In addition, gene expression profiles between cells with disparate chromosome numbers have been compared by mRNA-seq analysis.
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5:15 High-Throughput Screening of Transfection Efficiency of dTtaPS Reagent Library, and Its Application for Transient Production of Proteins in Micro Bioreactors

Harsh Jain, Ph.D., Visiting Associate, FDA

5:45 Close of Day


8:00 am Registration

8:00 BuzZ Sessions with Continental Breakfast

Protein therapeutics is a fast-growing global market. As the science improves, so does the complexity of the R&D organization. Ensuring product quality plus speed to market requires insights from stakeholders working across the stages of protein science R&D. Join experts representing this PepTalk pipeline, peers, and colleagues for an interactive roundtable discussion. Topics include highlights from the week’s presentations, new technologies and strategies, challenges, and future trends.

Table Moderator: Richard Altman, MS, Scientist, Protein Technologies, Amgen

Table Moderator: Bjørn Voldborg, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark


Technologies to Establish Efficient Production & Purification

9:00 Chairperson’s Remarks

Gabriel Rocklin, Ph.D., Senior Fellow, Biochemistry & Bioengineering, University of Washington

9:05 Platformization of Multi-Specific Protein Engineering I: From in silico Design and Bulk Modular Cloning to Automated Deconvolution of Variant Libraries

Joerg Birkenfeld, Ph.D., Section Head, High Throughput Biologics, R&D Biologics Research/Protein Therapeutics, Sanofi-Aventis Deutschland GmbH

The success rate to identify a multi-specific lead molecule with favorable drug-like properties increases with the number of variants tested. We report here the establishment of a novel, automated platform process for the fast generation of large panels of multi-specific variants (up to 10,000). Our high-throughput process integrates emerging cloning technologies with state-of-the-art automation and workflow supporting bioinformatics based on Genedata Biologics Database.

9:35 High-Throughput Methods for Protein Stability Prediction and Formulation Challenges Identification

Smita Raghava, Ph.D., Senior Scientist, Sterile Formulation Sciences, Merck & Co.

Successful development of biologics requires development of orthogonal tools to meet the challenge of rapidly and accurately assessing protein solution stability using limited material. This presentation will focus on combination of high-throughput technologies and assays for formulation and drug product development of biologics, such as monoclonal antibodies (mAbs) and mAb-based modalities. The overview of tools, their novel implementation, and relationship to commonly conducted “stability studies” will be further discussed using examples of high-throughput workflows, pre-formulation screening, and formulation development/optimization.

10:05 HTP Method for Affinity Determination in Complex Matrices by Solution Equilibrium Analysis Using Meso Scale Discovery Technology

Eilyn R. Lacy, Ph.D., Principal Scientist, Janssen BioTherapeutics (JBIO), Janssen Research & Development, LLC

10:35 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.

11:15 High-Throughput Automations and Optimizations for Improved Binder Generation and Validation

Jonas Schaefer, Ph.D., Head, High-Throughput Binder Selection Facility, Biochemistry, University of Zurich

While recombinant binder selection pipelines by now work in rather high-throughput, the screening of suitable affinity reagents and especially the validation of their essential features for the final applications is still laborious and time-intensive. To optimize the efficiency of these processes, we have improved already existing and developed novel methods to efficiently test candidates for their suitability, e.g., regarding their specificity.

11:45 High-Throughput Characterization of Hydrolytic Enzymes in Low Volume and Closed Systems

Nigel F. Reuel, Ph.D., Assistant Professor, Chemical and Biological Engineering, Iowa State University

Hydrolytic enzymes play a significant role in biologic and synthetic processes. The ability to better characterize these enzymes would enable shorter development times and better products. This talk will detail two recent developments for hydrolytic enzyme characterization: 1) a carbon nanotube-based optical sensor that allows for quantitative measurement in <20ul volumes, and 2) a resonant antenna sensor that passively transmits its response in the 1-100MHz range, enabling detection within closed, opaque systems.

12:15 pm Conference Wrap-Up

Richard Altman, MS, Scientist, Protein Technologies, Amgen

Haiyan Jiang, Ph.D., Principal Scientist, Biologics Research, Janssen BioTherapeutics, Janssen R&D

Diane Paskiet, MS, Director of Scientific Affairs, Scientific Affairs and Technical Services, West Pharmaceutical Services

Bjørn Voldborg, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark

Moritz von Stosch, Ph.D., Senior Manager, Fermentation, Technical R&D, GSK Vaccines


12:45 Close of Conference


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