Cambridge Healthtech Institute’s Fourth Annual
Optimizing Expression Platforms
Engineering Higher-Throughput Processes from Cells to Screens
January 12-13, 2017 | Hilton San Diego Bayfront | San Diego, CA
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 biotherapeutic
of interest. The rapidly increasing need for recombinant proteins necessitates further improvements in both technologies.
Cambridge Healthtech Institute’s Fourth 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.
THURSDAY, JANUARY 12
7:45 am Conference Registration and Morning Coffee
8:15 Chairperson’s Opening Remarks
Yves Durocher, Ph.D., Section Head, Mammalian Cell Expression - NRC Human Health Therapeutics Portfolio, National Research Council Canada
8:20 Strategies to Enable High-Throughput Recombinant Protein Production in Mammalian Cells for Preclinical Studies
Athena Wong, Ph.D., Senior Scientist & Senior Group Leader, Early Stage Cell Culture, Genentech
Transient transfections in HEK293 and CHO cells are used to rapidly generate proteins for discovery research and early development studies. Here we present our approaches to express microgram to multigram amounts of protein in automated 96 deep well plates
and bioreactors. To increase transfection productivity, we performed host cell engineering followed by process optimization. Results showed that modifying media components provides significant benefits towards increasing yield and/or modulating product
9:00 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
9:30 Development and Optimization of AAV hFIX Particles by Transient Transfection in an iCELLis Fixed Bed Bioreactor
Michael Meagher, Ph.D., Vice President, Therapeutics Production & Quality, St. Jude Children’s Research Hospital
The clinical demand for AAV requires a scalable high-capacity technology. The presentation describes the production of AAV8-hFIX using a 2 plasmid transient transfection of HEK293T/17 cells using a disposable fixed bed bioreactor, the iCELLis® Nano.
The iCELLis® can support as many as 2.5×108 cells/mL of fixed bed (1.9x106 cells/cm2). Optimizing culture and transfection parameters resulted in 9.0×1014 AAV8 viral particles per square
meter of fixed bed.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing
11:00 Optimizing CHO Transient Expression for Drug Discovery
Elizabeth Greene, MS, Scientist, Immune Modulation and Biotherapeutics Discovery (IMBD), Boehringer Ingelheim
Demand for expression of high-quality therapeutic antibodies and recombinant proteins is on a spiral rise, the process of which is laborious, time consuming and expensive. CHO cells have become a major workhorse for transient expression of recombinant
biologics. Transient CHO productivity is influenced by many factors, including clone design, vector backbone, codon usage, clone/host selection and process parameters operate in a matrix setting for the optimized production and yield of a functional
11:30 Strep-Tactin XT - A Superior Next-Generation System for Purification of Proteins & Assay Development
Dennis Niermeier, MSc, Scientist, IBA GmbH - Solutions for Life Sciences
The new third-generation Strep-tag® system is based on recently engineered Strep-Tactin®XT and Twin-Strep-tag®. Due to the affinity improved but still reversible binding of Strep-Tactin®XT to Twin-Strep-tag® in the low pM range, the
system is superior to other affinity purification systems and now also suitable for assay development.
11:45 Sponsored Presentation (Opportunity Available)
12:00 pm Session Break
12:15 Luncheon Presentation I: CHO & HEK293 New Screening Solutions, Tips and Tricks for Transient Transfection, and Stable Cell Lines
Sam Ellis, Vice President & Biochemist, Thomson Instrument Company
Evaluation of different transfection tools, product quality, and titer for both CHO and HEK293. CHO transient systems, reagents. Which system is best, and scalable? Titer vs. quality and repeatability for results? Data will be presented on techniques
and technology that allow for mimicking large-scale fermentation with non-controlled devices from 15mL-3L. All of these techniques will be given with existing case studies for technologies involving protein or antibody production, and can lead
to successful transfer from different protein groups.
12:45 Luncheon Presentation II (Sponsorship Opportunity Available)
1:15 Ice Cream Break in the Exhibit Hall with Poster Viewing
2:00 Chairperson’s Remarks
Richard Altman, MS, Scientist, Protein Technologies, Amgen
2:05 Chromatin Function Modifying Elements in an Industrial Antibody Production Platform
Mark Ellis, Principal Scientist, Protein Expression and Purification, UCB Pharma
The isolation of stably transfected cell lines for the manufacture of biotherapeutic protein products can be an arduous process. This frequently involves transgene amplification and maintenance over many generations. We assessed four chromatin function
modifying elements for their ability to negate chromatin insertion site position effects and their ability to maintain antibody expression. Stability analysis demonstrated that the reduction in expression was mitigated in the clones containing
2:35 Cell Line Profiling to Improve Productivity and Product Quality
Sohye Kang, Ph.D., Senior Scientist, Process Development, Amgen
Despite the same host cell origin, recombinant production cell lines often display phenotypic variability in regard to growth rate, cell size and metabolic profiles. These intrinsic, cell line-specific variations can affect productivity and product
quality. Technological advances in -omics and bioinformatics tools are providing unprecedented opportunity to probe the molecular systems that underlie various cellular phenotypes influencing quantity and quality of therapeutic protein production.
Findings from these investigations allow opportunities for process improvement.
3:05 Accelerating Biotherapeutic Development through Simultaneous High-Titer, CHO Transient Expression & Generation of High-Yield Stable Cell Lines Using Scalable Transfection
James Brady, Ph.D., Director, Technical Applications, MaxCyte, Inc.
3:35 Refreshment Break in the Exhibit Hall with Poster Viewing
4:15 Product Quality and Protein Expression in CHO Cells
Pauline Smidt, Process Development Scientist, Just Biotherapeutics
I discuss the evaluation of growth, titer and product quality in production CHO cell lines under various process conditions.
4:45 Rapid Production of Recombinant Proteins in CHO Cells Using Large-Scale Transfection or Stable Pools
Yves Durocher, Ph.D., Section Head, Mammalian Cell Expression - NRC Human Health Therapeutics Portfolio, National
Research Council Canada
We describe our CHO transient expression platform for rapid production of recombinant proteins. For more difficult-to-express proteins or proteins needed in large quantities, we also developed an inducible CHO pool platform that allows generation
of stable and scalable pools expressing high levels of monoclonal antibodies in two weeks post-transfection. Fc glycans present on monoclonal antibodies produced by transient transfection or stable pools are compared. The pools can also be used
to derive stable and high-expressing CHO clones for manufacturing therapeutic candidates.
5:15 Toolbox for Cell Line Development – Next-Generation Cell Line Development Technologies
Holger Laux, Ph.D., Fellow, Integrated Biologics Profiling, Technical Development NBE, Novartis
Chinese hamster ovary (CHO) cells are the most widely used host for large-scale production of recombinant therapeutic proteins exhibiting high productivities in the gram-per-liter range. A novel toolbox of vector elements, selection marker and novel
engineered CHO cell lines were developed which results in combination in significant increase of titer and improved product quality. We have evaluated novel vector elements and a new selection marker with a variety of antibody projects resulting
in an increase of titer. Furthermore, we have identified a key protein severely affecting the quality of non-antibody format therapeutic proteins. Subsequently the key protein was eliminated using Novartis propriety CHO cell line via novel targeted
gene disruption tools. This resulted in a superior CHO cell line. The combination of novel vector elements, novel cell lines and selection marker resulted in a significant increase of titers and improved product quality.
5:45 Close of Day
FRIDAY, JANUARY 13
8:00 am Conference Registration and Morning Coffee
8:30 Chairperson’s Remarks
Jonas V. Schaefer, Ph.D., Head, High-Throughput Binder Selection Facility, Biochemistry, University of Zurich
8:35 High-Throughput Methods for the Characterization of Relevant Protein Features
Jonas V. Schaefer, Ph.D., Head, High-Throughput Binder Selection Facility, Biochemistry, University
To optimize the efficiency of the laborious process of generating specific affinity reagents, we established a streamlined pipeline consisting of simultaneous selections against 94 targets and subsequent high-throughput screenings and validations.
This fast and efficient platform, allowing the reliable discovery of recombinant binders, requires the improvement of existing and the development of novel high-throughput methods which will be presented.
9:05 High-Throughput Biophysical and Biochemical Stability Screening for Early Stage Antibody Discovery
Yingda Xu, Ph.D., Associate Director, Protein Analytics, Adimab LLC
Problems in the development of antibodies can often be traced back to their intrinsic poor biophysical and biochemical stabilities. High-throughput screening assays are developed or adapted to fit in the scope of early discovery stage to filter out
candidates with poor properties.
9:35 High-Throughput Manufacturability Assessment of Complex Biologics
Zhenyu Gu, Ph.D., Development Scientist III, Early Assay Development, Alexion Pharmaceuticals
Bispecific/biparatopic antibodies, modified enzymes and fusion proteins have gained increasing popularity due to their unique therapeutic profiles. However, these molecules often pose significant challenges to manufacture as a result of their complex
designs. In this study, high-throughput mechanical/chemical stress relevant to manufacture process was coupled with high-throughput orthogonal characterization methods to screen these early stage molecules for their manufacturability, focusing
on solubility, aggregation, colloidal and thermal stability.
10:05 Coffee Break with a Poster Pavilion
11:00 Sequential Injection Capillary Electrophoresis for Bioprocess Monitoring
Rosanne Guijt, Ph.D., Alexander von Humboldt Fellow and Senior Lecturer, Australian Centre for Research
on Separation Science (ACROSS), University of Tasmania
Biological processes are naturally susceptible to variability because living cells consume substrates and produce metabolites and products in a dynamic way with variations in metabolic rate across short time intervals. This presentation explores the
potential of capillary electrophoresis (CE) for bioprocess monitoring. Using a novel injection strategy, this fully automated system offers high sample throughput, good temporal resolution and low sample consumption combined with robustness, sensitivity
and flexibility which provides a promising new platform for pharmacological and biotechnological studies.
11:30 High-Throughput Protein Analysis and Engineering Using Microcapillary Arrays
Spencer Alford, Ph.D., Protein Engineer, xCella Biosciences, Inc.
We developed a high-throughput screening platform that allows researchers to assay the functional activity of millions of protein variants, displayed on or secreted from cells. This talk describes several protein analysis and engineering applications
performed with this new technology platform.
1:15 Close of Conference