Cambridge Healthtech Institute’s 11th Annual

Optimizing Cell Line Development

Enhancing Expression

August  15-16 , 2019

 

Cell Line Development has reached a new plateau influenced by genomic research and insights, along with emerging technologies that are pushing cell engineering into an uncharted future. The “Optimizing Cell Line Development” conference brings together experts who are forging this new era. They will share how to best optimize codons, construct vectors, and how to select and engineer clones and host systems, while maintaining stability and consistency. The conference will also focus on genomic research for CHO and other systems, as well as glycoengineering, systems biology, assays, and pathway delineation.  In addition, challenges for introducing new technologies will be discussed, along with an overview of industrial trends and regulatory perspectives.

Final Agenda

Thursday, August 15

11:30 am Registration Open

12:15 pm Enjoy Lunch on Your Own

1:15 Refreshment Break in the Exhibit Hall with Last Chance for Poster Viewing (Sponsorship Opportunity Available)

KEYNOTE SESSION

1:55 Chairperson’s Remarks

Lorenz Mayr, PhD, Chief Technology Officer, GE Healthcare Life Sciences

2:00 KEYNOTE PRESENTATION: Cell Line Development in the Digital Age

Jennitte Stevens, PhD, Director, Process Development, Drug Substance Technologies, Amgen, Inc.

2:45 KEYNOTE PRESENTATION: Mammalian Synthetic Biology: Foundations and Application to Cell Line Engineering

Weiss_RonRon Weiss, PhD, Professor, Biological Engineering, Massachusetts Institute of Technology (MIT)

In this research, we appropriate from established engineering fields proven design principles such as abstraction, standardization, modularity, and computer aided design. But we also spend considerable effort towards understanding what makes synthetic biology different from all other existing engineering disciplines and discovering new design rules that are effective for the biological substrate. Building on this foundation, I will describe our recent application of synthetic biology tools and principles towards the improvement of cell line engineering and biomanufacturing.

3:30 Using siRNA for Cell Line Selection

Edward Eveleth, MS, CEO, HocusLocus, Inc.

Adding an siRNA to be co-transcribed with the gene of interest allows for a novel approach to cell line selection using transfected mRNA and magnetic isolation.

3:45 Sponsored Presentation (Opportunity Available)

4:00 Refreshment Break

ACCELERATING CELL LINE DEVELOPMENT TIMELINES

4:15 Novel Technologies to Accelerate Cell Line Development Timelines Close to the Biological Limit

Jostock_ThomasThomas Jostock, PhD, Senior Investigator II and Leading Scientist, Novartis Biologics Center, Novartis Pharma AG

This presentation will summarize how different novel technologies were implemented to enable aggressive acceleration of cell line development processes.


4:45 Platform Approaches Enabling Single-Cycle Cell Line Development

Linda Francullo, Principal Scientist, Drug Substance Development, Pfizer, Inc.

Historically, when time and resources where plentiful, the philosophy was if you can manipulate the CHO cells to express the product;  you could build enough bioreactors to supply Phase 1  and there was always time to make improvements prior to commercialization.  Recently, the paradigm has shifted to greatly reducing time and resources from Phase I to launch.  Advances in our cell line development platforms have enabled progress toward single cycle development reducing timelines and resources pre-clinical development to commercial launch. 

5:15 End of Day

Friday, August 16

7:30 am Registration Open

7:30 Small-Group Breakout Discussions with Continental Breakfast

This session provides the opportunity to discuss a focused topic with peers from around the world in an open, collegial setting. Select from the list of topics available and join the moderated discussion to share ideas, gain insights, establish collaborations or commiserate about persistent challenges.

DEVELOPING CHO-BASED CELL LINES

8:30 Chairperson’s Remarks

Thomas Jostock, PhD, Senior Investigator II and Leading Scientist, Novartis Biologics Center, Novartis Pharma AG

8:35 FEATURED PRESENTATION: Regulating Recombinant Protein Expression during CHO Pool Selection Increases Productivity

Durocher_YvesYves Durocher, PhD, Principal Research Officer, Human Health Therapeutics Research Center, 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 likely to fail surviving 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). This was observed with many r-proteins, including monoclonal antibodies, GPCRs and cytokines.

9:05 The Development of Advanced Synthetic Biology Tools to Accelerate CHO Cell Line Engineering

Cao_JicongJicong Cao, PhD, Postdoctoral Associate, Massachusetts Institute of Technology (MIT)

Traditional cell line engineering strategies using random gene integration and high-throughput colony screening is laborious and time-consuming. To address these issues, we have developed a recombinase-gene integration method for accelerated CHO cell line construction and enhanced expression. In addition, we have developed an ultrasensitive single-wall carbon nanotube-based protein detection method, which enables single-cell analysis of protein products secreted from various cell types.

9:35 Improved Single Cell Cloning Method for High Clonality Assurance

Ren Liu, PhD, Associate Principal Scientist, Merck & Co, Inc.

Chinese hamster ovary (CHO) cell is the work horse for biological drug manufacturing. Single cell cloning is a critical step in CHO production cell line development. We have developed a robust cloning method to improve CHO clone recovery and ensure clonality. The use of an animal-component free cloning medium can increase clone recovery by 50%. Meanwhile, combining single cell per well plating with clone imaging can ensure that clones are derived from single cell.

10:05 Networking Coffee Break

IMPROVING QUALITY

10:30 Complete Elimination of the Warburg Effect in CHO Cells

Hefzi_HoomanHooman Hefzi, PhD, Postdoctoral Researcher, Pediatrics, UC San Diego (UCSD)

Lactate has long been a problem in mammalian cell culture, requiring rigid bioprocess strategies to control. We show that simultaneous knockout of lactate dehydrogenase and ancillary regulatory enzymes reduces culture lactate to negligible levels. The resulting Warburg-null lines retain the same growth rate while gaining an extended proliferative period. The cells remain amenable to standard workflows for generating protein producing lines and maintain mature glycan structures. These cells are thus an attractive starting host cell line for industrial purposes.

11:00 An Artificial Golgi Reactor as an Alternative Method for Targeted Cell-Free Glycosylation

Makrydaki_ElliElli Makrydaki, MRes, MEng, Graduate Research and Teaching Assistant, Chemical Engineering, Imperial College London

We propose the design of an Artificial Golgi reactor for in vitro glycosylation of mAbs in a cell free system. By expressing selected glycosyltransferases and immobilising them on streptavidin coated beads we achieve sequential enzymatic reactions. By combining the benefits of a cell free system and the stability offered by immobilisation, the Artificial Golgi reactor can be an alternative production platform allowing enhanced product quality.

11:30 Process CO2 and Medium Ion Balance Control Strategy for Control of Galactosylation and Lactate Metabolism

Ahn_Woo_SukWoo Suk Ahn, PhD, Scientist Upstream, Process Science, Global Manufacturing Science and Technology (MSAT), Sanofi US

The product quality of biologics can be affected by process conditions, medium components, and cell metabolism. We demonstrate the modulation of product quality through controlling pCO2 and medium ion balances. The pCO2 control strategy and the prediction model by first principles lead to the switch of lactate metabolism from production to consumption mode, which was found to correlate to galactosylation.

12:00 pm Sponsored Presentation (Opportunity Available)

12:30 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:15 Session Break

IMPROVING PROCESSES

1:25 Chairperson’s Remarks

Elli Makrydaki, MS, Graduate Research and Teaching Assistant, Chemical Engineering, Imperial College London

1:30 Establishing a Robust HeLa Cell Line Screening Platform for Rapid, Scalable rAAV Production

Tiernan_AubreyAubrey R. Tiernan, PhD, Senior Scientist I and Head, Cell Line Development, Ultragenyx Gene Therapy

One challenge for cell line development in industry is to generate highly productive stable cell lines within the shortest time frame possible. This presentation will cover lessons learned over three years in establishing a robust, high throughput HeLa suspension screening platform to generate stable monoclonal producer cell lines suitable for Phase III clinical trial/commercial rAAV production.

2:00 The BEST of Both Worlds – Targeted Integration and Multiple Copies: How Can This Go Together for Improved Cell Line Development?

Anton Bauer, PhD, MBA, COO, R&D, The Antibody Lab GmbH

Targeted Hot Spot integration and multiplication of independent expression units – can this go together and even speed up cell line development? By targeting the Rosa26 Hot Spot in vitro we generated BAC-based expression vectors, which integrated in multiple copies into the CHO host cell chromatin and acted as independent expression units. This allowed us to adapt the selection process and developed long-term stable high-yield production cell lines at an unprecedented speed.

2:30 Driving Biological Discovery: An Expanding Toolkit for Affinity Proteomics

LaCava_JohnJohn LaCava, PhD, Research Assistant Professor, Laboratory of Cellular and Structural Biology, The Rockefeller University

It remains challenging to transfer intact physiological macromolecules from their native sources into suitably stabilizing in vitro environments. To address this, we developed an interactome capture platform that is akin to a crystallographic screen. The approach will be summarized in this talk, leveraging research vignettes. Our long-term objective is to enable the transfer of cognate healthy and pathogenic macromolecules from their in vivo milieus into test tubes, for biochemical, structural, and mechanistic studies.

3:00 Challenges of Stable Cell Line Development Using Horizon CHO GS Knock Out System

Qinghua Jenny Zhao, MSc, Associate Director, Process Development & Operation, Salubris Biotherapeutics

A stable cell line was developed for antibody production using a Horizon CHO GS knockout system through optimization of transfection, and culture medium during cloning steps. The electroporation transfection procedure with a Lonza system was developed based on different programs and buffers, cuvette size, host cell health, DNA quality and quantity. To increase transfection efficiency and protein expression, a FACS method was also introduced to verify targeted product expression and quality during clone selection.

3:30 Close of Conference




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