2020 Archived Content

Cambridge Healthtech Institute’s 7th Annual

Optimizing Expression Platforms

Employment and Development of Cells, Process, and Personnel to Efficiently Produce Biotherapeutics

January 23-24, 2020

Part of the Biotherapeutic Expression & Production pipeline

The utilization of engineered therapeutic proteins for basic research, clinical diagnostics, and therapy continues to expand. Consequently, protein expression laboratory managers and researchers face challenges for efficient expression, production, and purification, even while improving quantity and quality, plus minimizing time and cost. Transient protein production (TPP) has the advantage of speed and limited risk, while stable transfection – the longer and more complex process – has the advantage of being able to produce long-term expression of the biotherapeutic of interest. Cambridge Healthtech Institute’s 7th Annual Optimizing Expression Platforms conference convenes protein expression experts to share their day-to-day challenges in high-throughput production and purification labs and provide insights and solutions on meeting these challenges.

Final Agenda

THURSDAY, JANUARY 23

7:45 am Registration (Sapphire West Foyer) and Morning Coffee (Sapphire West & Aqua West Foyer)

8:10 Organizer’s Welcome Remarks

Mary Ann Brown, Executive Director, Conferences & Team Lead, PepTalk, Cambridge Healthtech Institute

8:15 Chairperson’s Opening Remarks

Xiaotian Zhong, PhD, Senior Principal Scientist, Lab Head, BioMedicine Design, Pfizer Worldwide R&D

KEYNOTE PRESENTATION

8:20 Antibody Cascade Continuous Paramagnetic Chromatography

John Kawooya, PhD, Director, Biologics Optimization, Discovery Research, Amgen, Inc.

“Cascade Continuous Paramagnetic Chromatography” (CCPC) purifies antibodies from crude cell cultures without centrifugation or filtration. In this process, Protein A paramagnetic beads are added to live cell cultures 18-24 hours prior to harvesting. This maximizes antibody binding, eliminates traditional cell removal and column loading steps. The contents are transferred into a single-use magnetizable tank for bead washing, antibody elution, viral inactivation, pH and ionic strength adjustment prior to SP-CEX polishing.

9:00 Scaling Up and Scaling Out: Pushing the Boundaries of Transient Protein Production

Wilkinson_Ian_PEX_Pipeline7 Ian Wilkinson, CSO, Absolute Antibody

Whilst transient yields have improved drastically in the last decade, scalable systems are time-consuming and costly to implement. Absolute Antibody has developed systems which scale up and scale out protein expression and purification, enabling the rapid and cost-effective production of milligram-to-gram quantities of large panels of proteins.

9:30 Developing Innovative Laboratory Products for Bioprocessing and Analytical Science

Mark Rehse, National Sales Manager, Thomson Instrument Company

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

11:00 Making a High Titer Transient System from a Stable CHO Platform

Harms_Peter Peter Harms, PhD, Principal Engineer, Cell Culture, Genentech

A high titer (500-1000 mg/L for antibodies) transient system was developed from the host cell and media of an in-house stable CHO platform. Development included screening of subclones and process optimization, but there were also a few surprises along the way. The final system is robust, scalable from 30 mL to 10 L, and provides product quality comparable to the original stable CHO platform.

11:30 Developing a Transient CHO Expression Platform with N-Glycan Compositions Consistent with Stable CHO

Zhong_Xiaotian Xiaotian Zhong, PhD, Senior Principal Scientist, Lab Head, BioMedicine Design, Pfizer Worldwide R&D

Developing a robust transient expression platform in CHO, the preferred host for many clinical and commercial products, offers a start-to-end quality alignment advantage for therapeutic protein discovery. This talk will present our recent efforts in developing a transient CHO platform to enable efficient sialylation of N-glycans. The talk will also describe new strategies in optimizing transient CHO system with N-glycan compositions consistent with those from stable CHO production.

12:00 pm NEW: A Universal Affinity-tag System: From Protein Purification to Analytic Applications

Karthaus_Dennis Dennis Karthaus, Director, Protein Products & Assays, IBA Lifesciences

New expression systems enable high titer production while simultaneously reducing the expression volume and time. Here we show how proteins can be easily and effectively purified from Expi supernatants using Strep-tag technology. Furthermore, Strep-Tactin®XT can be used in combination with BLI sensors for rapid measurement of protein concentrations and kinetics.

12:30 Session Break

12:40 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:10 Ice Cream Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

2:15 Chairperson’s Remarks

Yves Durocher, PhD, Research Officer, Bioprocess Engineering, National Research Council Canada

2:20 Reducing Recombinant Protein Expression during CHO Pool Selection Enhances Pool Productivity and the Frequency of High-Producing Cells

Durocher_Yves Yves Durocher, PhD, Research Officer, Bioprocess Engineering, National Research Council Canada

During the generation of stable cell lines, high-level expression of recombinant protein (r-protein) may impose a metabolic burden on the cells and many are not likely to survive the selection process. Using the cumate-inducible expression system, we show that selection in the “off-mode” allows the generation of stable pools with up to 3-fold higher productivity compared to selection in the “on-mode” (mimicking constitutive promoters). Pools selected in the “off-mode” contain a much higher proportion of high-producing cells.

2:50 Integrative Multi-Omics Data and Model-Driven Approaches Guide CHO Cell Line Development for Enhanced Bioprocessing

Lakshmanan_Meiyappan Meiyappan Lakshmanan, PhD, Research Scientist & Group Leader, Systems Biology, Bioprocessing Technology Institute, A*STAR

A systems approach based on high-throughput “-omics” profiling and mathematical modeling of CHO cells offers immense potential to become an indispensable tool in the biopharmaceutical industry for identifying key bottlenecks in the product yield and quality related pathways in a mechanistic manner. The knowledge obtained from such multi-omics data and model-guided systems approaches can further enable the next-generation cell line development.

3:20 Networking Refreshment Break (Sapphire West & Aqua West Foyer)

3:45 Optimization of NGS-Based Sequence Variant Analysis to Facilitate Cell Line Screening

Zhou_Yizhou Yizhou Zhou, PhD, Scientist II, Cell Line Development, Biogen

Reliable high-throughput analytic tools are critical for accelerating cell line development to push protein therapeutics into clinical trials. Sequence variants of the transgene can occur at multiple stages during the cell line development. Here we improved the sensitivity and robustness of an amplicon-based NGS assay, which fits into the aggressive timeline of cell line development and allows detection of very low levels of sequence variants for up to 40 clones.

4:15 Establishing a High-Throughput Protein Production Platform for Rapid Antigen Screening

Huang_Ying Ying Huang, MD, PhD, Lab Head, Vaccine Design and Characterization, Preclinical R&D, GSK Vaccines

Breakthrough technological innovations in the fields of recombinant protein expression and structural biology have provided new tools to accelerate vaccine design and optimization. Here we describe the establishment of a high-throughput antigen expression and purification platform using the Fluent® Liquid handling robot and the KingFisherTM Flex magnetic beads purification system compatible for antigen production in both bacterial and mammalian cells to rapidly screen hundreds of vaccine candidates from computational design.

4:45 PANEL DISCUSSION: Transient, Stable, or Both?

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.

Moderator:

Durocher_Yves Yves Durocher, PhD, Research Officer, Bioprocess Engineering, National Research Council Canada

Panelists:

Harms_Peter Peter Harms, PhD, Principal Engineer, Cell Culture, Genentech

Meiyappan Lakshmanan, PhD, Research Scientist & Group Leader, Systems Biology, Bioprocessing Technology Institute, A*STAR

Zhong_Xiaotian Xiaotian Zhong, PhD, Senior Principal Scientist, Lab Head, BioMedicine Design, Pfizer Worldwide R&D

Zhou_Yizhou Yizhou Zhou, PhD, Scientist II, Cell Line Development, Biogen

5:15 Close of Day

FRIDAY, JANUARY 24

8:00 am Registration (Sapphire West Foyer)

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.

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9:00 Chairperson’s Remarks

Richard Altman, MS, Field Application Scientist, Protein Expression, Biosciences Division, Life Sciences Solutions Group, Thermo Fisher Scientific

9:05 Running a Core Facility: How to Keep Everybody Happy

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

9:35 Balancing Platform Delivery, Innovation, and Job Satisfaction in the HT Protein Production Space

Kraft_Edward Edward Kraft, PhD, Senior Scientific Manager, Biomolecular Resources, Genentech

Working in high-throughput, non-antibody protein production labs represents unique challenges. Producing large batches of protein for diverse protein classes creates continuous challenges. I’ll discuss the approaches at Genentech to create a rewarding career experience that blends platform delivery, innovation and job satisfaction.

10:05 Solving Challenges to the Operation of a Protein Production Core Supporting Early Stage Drug Discovery Efforts

Esposito_Dominic Dominic Esposito, PhD, Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research

10:35 Networking Coffee Break (Sapphire West & Aqua West Foyer)

11:00 Delivering World-Leading Research Services to Drive Scientific Success

Somasundaram_Balaji Balaji Somasundaram, PhD, Strategy and Operations Manager, UQ Protein Expression Facility, The University of Queensland

Research service facilities are becoming a vital component of the modern collaborative research environment, where researchers use the technical expertise and experience of the facility to advance their research. This presentation will cover the following key focus areas for a protein research service facility to be integrated as a valued partner in research programs: 1) Enhancing operational excellence; 2) Building strategic partnerships; and 3) Workforce development.

11:30 Balancing Innovation, Efficiency, and Quality in Protein Production

Williamson_Jessica Jessica Williamson, PhD, Protein Production Lead, UCB Biosciences

Running a protein production group in drug discovery requires a lot of organization. We’re all engaged in the actual research, but from managing resources and meeting deadlines to maintaining quality and maximizing yield, it’s a challenging balancing act. At UCB Biosciences, we have years of experience as a gene-to-structure pipeline and we are applying the lessons we’ve learned to increase efficiency, embrace innovation, and maintain high quality.

12:00 pm CLOSING PANEL DISCUSSION: Protein Production Lab Challenges: Methodologies, Strategies, and the Art of Managing Multiple Projects

There are many challenges in operating protein production labs. This panel focuses on the following topics: initiating projects, basic expression and purification systems, pros and cons for each system, screening platforms, troubleshooting and how much time should be spent on each system before moving to the next option. In addition to “hands-on” tips, we touch upon strategies on how to manage multiple “top priority” projects.