Cambridge Healthtech Institute’s 3rd Annual

Cell Line Development to Protein Expression

From Gene to Industrial Use

25-26 March 2020

 

The foundations of protein production will be explored in Cambridge Healthtech Institute’s Cell Line Development to Protein Expression conference. What are the steps needed to develop a productive cell line that meets project goals? Each protein and each expression system is different, so which is best to use? This conference will examine the steps that are necessary to establish productive cell lines that ensure robust expression to support the industry’s growing needs.

Final Agenda

Wednesday, 25 March

10:30 Registration

10:30 Coffee Break in the Exhibit Hall with Poster Viewing

PLENARY SESSION:

NEXT-GENERATION PROCESSES AND PRODUCTS

11:15 Chairperson’s Remarks

Jarka Glassey, PhD, Professor, Chemical Engineering, Engineering, Newcastle University

11:20 Current Opportunities and Challenges in Biotherapeutic CMC

Schumacher_RalfRalf Schumacher, PhD, Global Head, Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG

As biologic pipelines continue to grow and diversify, there is an increasing need to standardize, automate, and find efficiencies along the entire value chain. This presentation will discuss the current challenges and opportunities in biotherapeutic CMC and the impact new modalities are having on upstream and downstream processing, analytics, and formulation. The advantages of predictive process parameters in early-stage development and digital development concepts to speed up the CMC development will also be discussed.

11:50 Gene Therapy Manufacturing and Technical Development

Blumenthal_DianeDiane Blumenthal, PhD, Head, Technical Development, Spark Therapeutics, Inc.

In the past few years, several cell and gene therapy products have gained regulatory approval in the US and EU with many more in the pipeline. Manufacturers of cell and gene therapy products must tackle technological challenges under the pressure of short timelines resulting from streamlined clinical development. This presentation will focus on the key technical development challenges facing the industry as product development programs move into the later stages of process development and scale-up, process performance qualification, and ultimately, commercialization.

12:20 Session Break

12:30 Bridging Luncheon
          Presentation to be AnnouncedMERCK(1)

13:00 Session Break

REFINING CHO/MAMMALIAN ENGINEERING

13:45 Chairperson’s Opening Remarks

Philip Probert, PhD, Senior Scientist, Upstream, Centre for Process Innovation, Ltd.


13:50 KEYNOTE PRESENTATION:
Engineering of Multiple Metabolic Pathways Mediates Improved Productivity in CHO Cells

Nicolas P. Mermod, PhD, Professor & Director, Biology & Medicine, University of Lausanne

We uncovered numerous cellular genes that are overexpressed in CHO cell lines that produce high levels of therapeutic proteins. While some were the consequence of heterologous protein overexpression, others appeared to limit the expression and secretion of heterologous proteins. This was assessed by overexpressing these genes, resulting in increased expression of various therapeutic proteins. Interestingly, several CHO cell metabolic activities were simultaneously limiting in pathways as diverse as cell signaling, response to cellular stress, cytoskeleton organization, and lipid metabolism.

14:20 Studying the Impact of Genetic Alterations Using a Targeted Integration CHO Host Cell Line

Trautwein_MarkMark Trautwein, Dr. rer. nat., Senior Scientist, Research & Development, Pharmaceuticals, Expression Technologies, Bayer AG

Both the chromosomal environment of the integration site, as well as the genetic elements of a transgene expression cassette contribute to the degree of high and stable transgene expression. We have used targeted integration host cell lines for evaluation of different genetic elements of the transgene construct, as well as for CRISPR/Cas9-based implementation of targeted (epi)genetic modifications. This approach facilitates optimization of product-specific expression configurations.

14:50 Glycoengineering CHO Cell Lines for the Production of Hard-to-Produce Glycoproteins

Voldborg_BjoernBjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark (DTU)

Using our high-throughput cell-line engineering platform, we have engineered a panel of CHO cells able to produce proteins with specifically designed glycoprofiles. We have used these cells to produce therapeutic proteins that have previously not been possible to produce in CHO cells. This approach can be used to produce therapeutic proteins with better biological properties, such as increased half-life, improved activity, etc.

15:20 Presentation to be AnnouncedIprasense

15:35 Sponsored Presentation (Opportunity Available)

15:50 Refreshment Break in the Exhibit Hall with Poster Viewing

16:25 CHO Cells Characterisation and Manipulation for Increased Recombinant Protein Production

Pekle_EvaEva Pekle, PhD, Associate Scientist, Process Development, Achilles Therapeutics, Ltd.

Biopharmaceutical technology development aims to produce recombinant proteins with desired product quality at commercial quantities. To increase productivity in CHO cells, correlative links between cell attributes and titre were investigated, and a cell attribute was found to be predictive of productivity. Cell sorting based on that cell attribute via FACS-generated pools/cell lines with increased productivity compared to unsorted cells. A new method to enrich high expressers has been developed.

16:55 Rapid Selection of CHO Clones Secreting Recombinant Antibodies

Devriendt_BertBert Devriendt, PhD, Scientist, Virology, Parasitology, Immunology, Physiology, Ghent University

To enable large-scale recombinant antibody production, a high-producer cell line is essential. Selecting such a cell line is, however, time-consuming and labor-intensive. By combining the design of a tricistronic vector expressing GFP and both antibody chains, separated by a GT2A sequence, with single-cell sorting and automated image analysis, a CHO cell line was rapidly selected producing high amounts of recombinant antibodies, which showed minimal degradation.

17:25 Precise Genome Engineering of Mammalian Cells for Antibody Expression and Screening

Parola_CristinaCristina Parola, PhD, Scientist, Biologics Research, Sanofi-Aventis Recherche & Développement, Sanofi

By taking advantage of CRISPR-Cas9 genome editing, we have developed a novel mammalian cell system for the expression of full-length antibodies. The Plug-and-(Dis)play (PnP) workflow included the initial generation of a platform cell line in which a novel specificity is introduced by means of a synthetic antibody. Subsequently, we optimized HDR efficiency to render the system amenable to antibody discovery and engineering; specifically, we were able to display libraries for the discovery of antibodies from immunized animals, and to affinity mature a known antibody by screening a library of random mutagenesis variants.

17:55 End of Day

18:00 Dinner Short Course Registration


Recommended Dinner Short Course*

18:30 - 21:00 SC5: Saving Time in Process Development with Next-Generation Methods: PAT, Hybrid Modeling, Process Simulation, mDoE, and iDoE

Instructors: Moritz von Stosch, PhD, Senior Manager, Drug Substance, Technical R&D, GSK Vaccines

Mark Dürkop, PhD, Project Leader, Biotechnology, University of Natural Resources and Life Sciences, Vienna (BOKU); Co-Founder and CEO, Novasign, Inc.

*Separate registration required.

Thursday, 26 March

8:00 Registration and Morning Coffee

STRATEGIES TO BOOST PRODUCTIVITY

8:25 Chairperson’s Remarks

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

8:30 FEATURED PRESENTATION:
Manipulating the Epitranscriptome as a Means to Impact Recombinant Protein Production in Mammalian Cells

Barron_NiallNiall Barron, PhD, Professor, Principal Investigator, Cell Engineering, National Institute for Bioprocessing Research and Training (NIBRT)

Epigenetic modifications to the nucleotides in RNA species have generated considerable interest recently. The role of methylation in particular, including characterising the proteins that add (writers), remove (erasers), and interpret (readers) this epigenetic mark, is considered. This talk will consider its potential as an enhancer of mRNA translation and how this mechanism might be applied to improving recombinant protein expression systems.

9:00 Strategies for Production and Characterization of Multi-Protein Complexes: Top-Down vs. Bottom-Up Strategies

Poterszman_ArnaudArnaud Poterszman, PhD, Research Director, Integrated Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC)

Although technologies for production of complexes have been tremendously improved, sample preparation is still a major bottleneck for structural biology and functional analysis both in academia and industry. Using the 10 subunits transcription/DNA repair factor TFIIH, I will illustrate reconstitution of multisubunit complexes using the baculovirus expression system and characterization of endogenous assemblies from CRISPR/Cas9-edited cell lines.

9:30 Speeding Up Cell-Line Development for the Expression of mAbs and Bispecific Antibody Formats

Martin Bertschinger, PhD, Deputy Director, Cell Sciences, Ichnos Sciences, Inc.

In many cases, the cell-line development (CLD) process is on the critical path of projects towards IND filings. In order to shorten the time required to start clinical trials, there is a strong interest in speeding up the duration of the CLD process. Ichnos Sciences has evaluated the implementation of a single-cell deposition device in the CLD process. This presentation will provide a critical review of this technology with regards to the desired shortening of timelines, as well as clonal derivation (“monoclonality”), productivity, product quality, and expression stability of the resulting cell populations.

10:00 GlycoExpress® - An Alternative Host for Difficult to Express ProteinsGlycotope

Goralczyk_VickyVicky Goralczyk, PhD, Director, Cell Line and Bioprocess Development, R&D, Glycotope GmbH

Bio-pharma development is shifting rapidly from classical IgG molecules to more challenging complex biopharmaceuticals like bispecifics or non-IgG molecules. With CHO being a good production cell line for IgG molecules, they might fail to produce more challenging candidates. The GlycoExpress® (GEX®) system represents an ideal alternative for the production these difficult to express protein molecules and will provide case studies which demonstrate the superiority in productivity and product quality vs CHO cell expression.

10:30 Coffee Break in the Exhibit Hall. Last Chance for Poster Viewing.

IMPROVING CELL LINE DEVELOPMENT

11:15 How to Be Good at Being Single? Evaluation of VIPS Single-Cell Printer for Generation of Stable Cell Lines

Kozejova_GabrielaGabriela Kozejova, MSc, Senior Scientist, Protein and Cellular Sciences, R&D Platform Technology & Science, GSK Medicines Research Centre

Stable cell-line generation is a time-consuming and labour-intensive process. We report the impact of implementing systems such as a Verified In-situ Plate Seeding system (VIPS™) linked with the Cell Metric imager. This resulted in an increase in throughput, reduced timelines, and provided clonality assurance for our stable cell-line processes.

11:45 A Generic Process for the Production of κλ Body Mixtures from a Single Cell Line

Poitevin_YvesYves Poitevin, MSc, Head, Bioprocessing Unit, Light Chain Bioscience (a brand of NovImmune SA)

One approach that has recently gained interest is the combination of antibodies included into a single dosage as a “multi-epitope-targeting” strategy mostly for oncology and infectious disease indications. By exploiting our unique technology platform to generate fully human bispecific antibodies (κλ bodies), a robust process compatible with industrial-scale manufacturing for the generation of stable monoclonal cell lines producing mixtures of mono- and bispecific antibodies will be described.

12:15 Do’s and Don’ts in Translating Cell-Line Research from Academia to Biotech

Danieli_TsafiTsafi Danieli, PhD, Head, Protein Expression Core Facility, Wolfson Center for Applied Structural Biology, The Hebrew University of Jerusalem

There are often basic differences in establishing producing cell lines and analyzing protein quality between academia and industry. How do we set ground rules for both basic research and early-stage companies to improve translation? The talk will focus on trying to set basic rules and examples of do’s and don’ts for improving tech transfer and facilitate the translation of academic language to industry.

12:45 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

13:30 Session Break

TARGETED APPROACHES TO ENGINEERING CELL LINES

13:45 Chairperson’s Remarks

Tsafi Danieli, PhD, Head, Protein Expression Core Facility, Wolfson Center for Applied Structural Biology, The Hebrew University of Jerusalem

13:50 Continuous Production with E. coli – USP Concepts and Strategies

Striedner_GeraldGerald Striedner, PhD, Associate Professor, Biotechnology, University of Natural Resources and Life Sciences, Vienna (BOKU)

Genetic stability of E. coli expression systems represents the major barrier in context with development of continuous production processes. In this talk, data from successful solutions, comprising host-cell engineering (genome-integrated plasmid-free systems, growth-decoupled systems), and USP process design strategies (mult-istage cultivation) will be presented and aspects for further improvements will be discussed.

14:20 Producing Stable Cell Lines Generated by Lentiviral Vectors

Chen_MarkMark Mao Xiang Chen, PhD, Manager, Cell Line Development and Cell & Gene Therapy, GlaxoSmithKline


14:50 Glycoengineering of the Human Embryonic Kidney FreeStyle 293-F Cell Line towards Improved Bioavailablity of Recombinant Coagulation Factor VII

Uhler_RicoRico Uhler, MSc, Scientist, Cell Line Development, Octapharma Biopharmaceuticals GmbH

In order to increase the bioavailability of coagulation Factor VII, the N-glycosylation machinery of the human embryonic kidney, FreeStyle 293-F (HEK 293 F), cell line was engineered using CRISPR/Cas9. N-acetylgalactosamine transferase knock-outs led to the replacement of N-acetylgalactosamine with galactose and increased sialylation. Knock-in of the sialic acid transferase, ST6Gal1, further increased sialylation of Factor VII and greatly reduced glycosylation-related heterogeneity of Factor VII molecules. Factor VII produced in the engineered cell lines showed reduced asialoglycoprotein receptor binding in vitro and increased bioavailability in vivo.

15:20 Close of Summit

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