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
THURSDAY, JANUARY 11
7:45 am Registration and Morning Coffee
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 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 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 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
Christopher 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 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 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 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 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 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 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 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.
View the Interview
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
FRIDAY, JANUARY 12
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
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