Optimising Expression Platforms banner

- 表现优化 -

The utilisation of engineered therapeutic proteins (recombinant proteins/recombinant antibodies) for basic research, clinical diagnostics and therapy continues to expand. Consequently, the efficient expression and production of these valuable biomolecules face challenges in improving their quantity and quality while minimising time and cost. To meet these demands, an increasing variety of recombinant production platforms called “cell factories” are being developed. Unfortunately, there is no “universal” production system which can guarantee high yields, particularly as every protein itself causes its own issues in terms of expression and production.   

Through case studies, the Optimising Expression Platforms conference offers comparisons, evaluations and solutions that enable protein engineers to efficiently express the therapeutic protein of their choice. 


Final Agenda

WEDNESDAY 14 NOVEMBER

07:45 Registration and Morning Coffee

TRANSIENT EXPRESSION AND PRODUCTION

08:30 Chairperson’s Remarks

Richard Altman, MS, Scientist, Protein Technologies, Amgen


08:35 KEYNOTE PRESENTATION: Transient Protein (Gene) Expression: From R&D towards Pharmaceutical Manufacturing

Florian M. Wurm, Dr. rer. nat., Professor Emeritus, Swiss Federal Institute of Technology Lausanne (EPFL); Founder, Chairman, ExcellGene SA

Transient protein (gene) expression (TGE) delivers products in days. HEK-293 cell TGE helped to identify a “better than nature” thrombolytic resulting in the approval of (TNKase “Tenecteplase”). Scale-up R&D resulted in the first 100 Liter–scale production in the late 1990s. Recent progress in transient protein expression, including viral vector production, delivers grams and tens of grams of high-quality protein/virus vector for preclinical research and, for clinical use soon?

09:05 Stable versus Transient Gene Expression: A Case Study on Antibody Glycosylation

Cleo Kontoravdi, PhD, Reader, Biosystems Engineering, Department of Chemical Engineering, Imperial College London

In this case study, we explored the differences in CHO cell metabolism, antibody productivity and N-linked glycosylation between stable and transient gene expression at physiological temperature and under mild hypothermia. For each system, we identified bottlenecks for improving antibody quality and have attempted to address them using cell and process engineering.

09:35 How Do We Assemble an Effective and Efficient Protein Production Toolbox?

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.

10:05 Selexis’ SUREscan and SUREsignature: Technologies for Assessment of Cell Line Integrity and Clonality

Igor Fisch, CEO, Selexis SA

Selexis’ SUREscan utilizes NGS technologies and proprietary bioinformatics to comprehensively assess the genomic architecture of Selexis-generated cell lines. SUREsignature is a unique, genome-wide collection of genetic markers that establishes cell line clonality to a probability significance not previously achievable and determines cell line-specific barcodes for better master cell bank quality control. These technologies minimize risk and accelerate biologics development by allowing for optimal clone selection and monitoring.

10:35 Coffee Break in the Exhibit Hall with Poster Viewing

11:15 Tracking the Evolution of Transiently Transfected Individual Cells in a Microfluidic Platform

Sébastien Sart, PhD, Research Associate, Genome and Genetics – Laboratory of Physical Microfluidics and Bioengineering, Institut Pasteur; Laboratoire d’Hydrodynamique, École Polytechnique

A droplet microfluidic platform was used to assess the heterogeneities of CHO-S transiently transfected with cationic liposomes. A single cell analysis of the GFP production kinetics revealed the presence of a subpopulation producing higher levels of GFP, which was dependent on the cell size, the charge and the amount of plasmid DNA carried by the lipoplexes. This study demonstrates the potential of time resolved single cell measurements to explain population dynamics.

11:45 Discovery of Novel Enhancers for Antibody Expression by Transcriptomics-Based Pathway Analysis

Markus Neubauer, PhD, Head, Cell Culture Research, Pharma Research & Development, Roche Innovation Center Munich

Expression of antibodies by transient transfection (TT) represents a widely used technology in basic research and drug discovery. Small molecule enhancers of expression are applied to maximize protein yield. This study demonstrates a successful strategy for identification of expression enhancers using comparative transcriptomics in conjunction with bioinformatics pathway analysis. Both pathways affecting antibody expression and molecules enhancing antibody expression have been discovered. This strategy may also be relevant for development and optimization of bioprocesses beyond TT.

12:15 High-Throughput Antigen and Antibody Production at the IPI

James Love, PhD, COO, Institute for Protein Innovation, Harvard Institutes of Medicine

In order to generate open-source monoclonal antibodies against every extracellular and secreted protein in humans, we have developed expression platforms capable of generating high-quality antigens and antibodies in HT format. Optimized transient transfection is performed via automated processes at 1ml and 30ml scales, and semi-automated for larger scales in HEK and CHO cells. Novel DNA preparation, protein purification and characterization platforms have been implemented to support the expression pipeline.

12:45 Scaling Up and Scaling Out: Pushing the Boundaries of Transient Protein Production

Ian Wilkinson, CSO, Research and Development, Absolute Antibody Ltd.

Whilst transient yields have improved drastically in the last decade, scalable systems are time-consuming and expensive 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.

13:00 Value Adding Microbial-Based Solutions for the GMP-Production of Recombinant Proteins

Nicole Peuker, PhD, Principal Expert USP Development, BioProcess Development, Wacker Biotech GmbH

Wacker Biotech, known as the microbial CDMO, handles several GMP production sites in Europe with capacities to deliver multiple hundred grams of drug substance per batch. We will present case studies for our innovative and cost-saving E. coli technologies for the production of difficult-to-make biopharmaceuticals.

13:15 Luncheon Presentation I to be Announced

13:45 Luncheon Presentation II: Accelerating Timelines by Integrating Cell Line Development and Manufacturing Programs

Simon Keen, Head, Cell Line Development, Biology, Abzena

14:15 Session Break

TOOLS FOR INCREASING EXPRESSION AND PRODUCTION

14:30 Chairperson’s Remarks

Henry C. Chiou, PhD, Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific

14:35 Synonymous Codon Selection for Enhanced Yield of Functional Proteins

Patricia L. Clark, PhD, O’Hara Professor of Chemistry & Biochemistry; Concurrent Professor of Chemical & Biomolecular Engineering, University of Notre Dame

We have developed a sensitive system to detect effects of synonymous codon substitutions on the co-translational folding of proteins expressed in E. coli, coupling the success of folding to E. coli fitness. We find that position-specific synonymous codon changes can have dramatic effects on folding yield, particularly at those positions that correspond to sub-domain “motif” structures.

15:05 Development of Myceliophthora thermophila into a Highly Productive Biologics Production Host

Marika Vitikainen, PhD, Senior Scientist, Industrial Biotechnology and Food Solutions, VTT Technical Research Centre of Finland Ltd.

Myceliophthora thermophila is well known as a hyper-productive platform for industrial enzymes. We are further developing a high-level, low-cost production host for biotherapeutics, such as antibodies, Fc-fusion proteins and vaccines. We have adapted it by modifying regulatory genes, reducing secreted proteases and altering glycosylation pathways needed for adding mammalian glycoforms. As a result, this system has reached antibody productivity levels over 1.7 g/L/day, much higher than current best-in-class platforms.

15:35 Refreshment Break in the Exhibit Hall with Poster Viewing

16:15 Identification and Visualization of Production Bottlenecks in CHO Cells

Kerstin Otte, PhD, Professor, Biotechnology, Biberach University of Applied Sciences

With the advance of complex format proteins, mammalian expression systems often show low performance. Determining factors may be the accumulation or haltering of heterologous proteins within the different cellular compartments, thus disturbing transport or secretion. We established a streamlined microscopy-based methodology for CHO production cells investigating the distribution of difficult-to-express proteins within organelles of the secretory pathway to enable identification of rate-limiting steps. This method was adapted for automated detection of production bottlenecks during industrial cell line development processes.

16:45 Streamlining Monoclonal CHO Cell Line Generation Using Droplet Microfluidics

James White, Research Scientist, Protein Sciences, UCB Biopharma

To increase the efficiency and speed of selecting high productivity monoclonal stable CHO cell lines, we have evaluated a droplet microfluidics device. This device screens for secreted antibody from millions of single CHO cells in their own pL volume droplets and to deposit the highest producers at a single cell per well. Our data demonstrates that high productivity clones similar to those generated in our standard processes can be selected, but with reduced resource needs and time to therapeutic clone selection.

17:15 PANEL DISCUSSION: Transient, Stable, or Both?

Moderator:

Henry C. Chiou, PhD, Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific

Panelists:

Richard Altman, MS, Scientist, Protein Technologies, Amgen

Cleo Kontoravdi, PhD, Reader, Biosystems Engineering, Department of Chemical Engineering, Imperial College London

James Love, PhD, COO, Institute for Protein Innovation, Harvard Institutes of Medicine

Markus Neubauer, PhD, Head, Cell Culture Research, Pharma Research & Development, Roche Innovation Center Munich

Florian M. Wurm, Dr. rer. nat., Professor Emeritus, Swiss Federal Institute of Technology Lausanne (EPFL); Founder, Chairman, ExcellGene SA

17:45 Networking Reception in the Exhibit Hall with Poster Viewing

18:45 Problem-Solving Breakout Discussions

19:45 End of Day

THURSDAY 15 NOVEMBER

08:00 Registration and Morning Coffee

DIFFICULT-TO-EXPRESS PROTEINS

08:30 Chairperson’s Remarks

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

08:35 FEATURED PRESENTATION: High-Throughput Expression and Screening of Human Integral Membrane Proteins

Nicola Burgess-Brown, PhD, Principal Investigator, Biotechnology, Structural Genomics Consortium (SGC), University of Oxford

The SGC promotes research advancement through our open access policy, and in the absence of IP. Globally, we have solved more than 2000 human protein structures and 10 novel integral membrane proteins (IMPs). Although we have made a significant contribution to structural biology and protein production for functional studies, IMPs and protein-protein complexes still remain a challenge to produce. Here, I present our established approaches for eukaryotic expression and screening IMPs using baculovirus/insect cells and BacMam technology.

09:05 Challenging Molecules in Biopharmaceutical Development: Innovative Tools for Cell Line Development

Martin Gamer, PhD, Associate Director, Early Stage Bioprocess Development, Boehringer Ingelheim Pharma GmbH & Co. KG

The increasing number of engineered, often antibody-derived molecule formats entering into biopharmaceutical development poses significant challenges on the generation of high-yielding CHO cell factories. My talk highlights the most recent advances at Boehringer Ingelheim to improve cell line development of DTE proteins. Our toolbox comprises in silico methods to assess molecule developability leading to tailored development, a rationally designed novel host cell line ensuring high performances, robustness and scalability as well as innovative genetic elements and screening tools to select for outstanding CHO production cell lines.

09:35 Producing a Glycosylating Escherichia coli Cell Factory

Phillip Wright, PhD, Faculty Pro-Vice-Chancellor, Faculty of Science, Agriculture & Engineering, Newcastle University

10:05 Presentation to be Announced

10:35 Coffee Break in the Exhibit Hall with Poster Viewing

11:15 Making and Testing the Human Secretome

Rick Davies, PhD, Associate Director, Discovery Biology, AstraZeneca Pharmaceuticals

The objective of the Human Secretome Project is to produce and screen all human secreted proteins to unlock biology leading to new hypotheses and target discovery. Over 1,000 secreted proteins have been expressed and purified using a mammalian expression system and screened in a number of cell-based phenotypic assays. The results of this work have revealed differential activities of protein family members in different biological contexts and provided some learning on successes and failures of recombinant expression of secreted proteins.

11:45 FEATURED PRESENTATION: Engineering CHO Cells for Production of Hard-to-Produce Proteins

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

Through extensive systems biology-based cell line engineering, we have engineered CHO cells for the production of therapeutically relevant proteins that were previously not possible to produce using CHO cells.

12:15 Luncheon Presentation I: Streamlined Discovery and Production of Therapeutic Antibodies

Meelis Kadaja, PhD, MBA, Director, Business Development, Icosagen Cell Factory

We take advantage of the universal HybriFree antibody discovery engine to efficiently discover therapeutic antibodies by direct cloning from B-cells of immunized rabbit, chicken, human, or dog. HybriFree method is further powered by our patented QMCF expression platform to produce premium-quality recombinant protein antigens, and antibodies cost-effectively for preclinical research (including afucosylated antibodies for enhanced ADCC). Technologies and case studies will be presented and discussed.

12:45 Luncheon Presentation II (Sponsorship Opportunity Available)

13:15 Dessert Break in the Exhibit Hall with Poster Viewing

14:00 End of Optimising Expression Platforms

17:00 Dinner Short Course Registration*

17:3020:30 Dinner Short Courses


Recommended Short Course*

SC9: Optimising Protein Purification Strategies in Advance: Getting Your Plan Right

David O’Connell, PhD, Lecturer, Biotherapeutics, Biomolecular & Biomedical Science, University College Dublin

This course addresses creating an effective strategy for purifying protein before beginning a purification project. What are key considerations before you launch an expression campaign? Which host should you select and why (Bacterial/Insect/Mammalian)? We will examine ways to reduce complexity in your strategy in order to efficiently increase productivity. We will also discuss how to establish redundant steps that support and guide protein purification.

*Separate registration required.

* 活动内容有可能不事先告知作更动及调整。

Choose your language
Chinese
Japanese
Korean
English