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第16回年次会議「発現プラットフォームの最適化」

基礎研究、標的開発、臨床診断、治療における遺伝子組換えタンパク質の利用は拡大し続けています。その結果、これらの貴重なバイオ分子の効率的な発現や生産は、時間とコストを最小限に抑えながら、量と質を向上させるという課題に直面しています。これらの需要に応えるため、「セルファクトリー」と呼ばれる、多様な組換え生産プラットフォームが開発されています。残念ながら、タンパク質の発現や生産にはそれぞれ独自の問題があり、高収量を保証できる「ユニバーサル」な生産システムはありません。Cambridge Healthtech Instituteの「発現プラットフォームの最適化」会議では、ケーススタディを通じて、タンパク質発現の研究者が、選択した治療用タンパク質を効率的に発現させるための比較・評価・ソリューションを提供します。

Recommended Short Course*
Monday, 13 November, 14:00 - 17:00
SC4: The Use and Optimization of Eukaryotic Expression Systems to Support Therapeutic Generation and Structural Biology
*Separate registration required. See short courses page for details. All short courses take place in-person only.

11月15日(水)

Registration Open and Morning Coffee07:30

OVERCOMING EXPRESSION AND PRODUCTION CHALLENGES OF DIFFICULT-TO-EXPRESS PROTEINS
発現が難しいタンパク質における発現・生産の課題の克服

08:25

Chairperson's Opening Remarks

Ana Sofia Coroadinha, PhD, Lab Head, Health & Pharma Division, Animal Cell Technology Unit Cell Line Development and Molecular Biotechnology Lab, IBET

08:30

Development of Specialized Bacterial Strains for High-Level Production of Recombinant Membrane Proteins

Georgios Skretas, PhD, Director, Institute for Bio-innovation, Biomedical Sciences Research Center "Alexander Fleming;" Founder & CEO, ResQ Biotech

We will describe the development of first- and second-generation E. coli SuptoxD and SuptoxR, two specialized strains for high-level recombinant membrane protein (MP) production. These engineered strains can: (1) suppress the toxicity that frequently accompanies MP overexpression, thus enabling enhanced levels of final bacterial biomass; and (2) markedly increase the cellular accumulation of membrane-embedded protein. Combined, these two positive effects result in dramatically enhanced volumetric yields for various prokaryotic and eukaryotic recombinant MPs.

08:50

Membrane Protein Production Using Insect Cells

Alice Rothnie, DPhil, Senior Lecturer, Biochemistry, Aston University

Membrane proteins play important roles in cell signaling, the influx and efflux of nutrients and metabolites, and many are potential drug targets. For their structural and functional analysis, high yields of correctly folded and modified protein are needed. Multidrug resistance protein 4 (MRP4/ABCC4) is a multi-substrate primary transporter that has been expressed using both recombinant baculovirus infection of Sf9 insect cells, and baculovirus-mediated transduction of Freestyle HEK cells.

09:10

Lactococcus lactis, a Promising Cell Factory to Functionally Express Membrane Proteins

Annie Frelet-Barrand, PhD, Researcher, MN2S, Institut FEMTO-ST

Membrane proteins (MPs), important drug targets, display crucial functions in organisms but their study remains difficult (hydrophobicity and low abundance). Their overexpression is mandatory for structural and functional characterizations but this strategy could encounter obstacles. Using the NICE system and the L. lactis strains, in the last twenty years, more than 100 MPs were expressed allowing either their functional and/or structural characterization. Recently, one eukaryotic membrane protein was expressed at a high-expression yield and allowed the formation of intracellular vesicles. In conclusion, L. lactis represents an interesting system for expression of MPs.

09:30 Talk Title to be Announced

Speaker to be Announced

09:45 Fyonibio's Versatile Cell Line Expression Platforms for the Development of Complex Molecules

Lena Thoring, Director, Cell line and Bioprocess Development, FyoniBio GmbH

Here FyoniBio presents its versatile highly productive expression platforms, the mammalian host cell systems CHOnamite®, for e.g. bi-specific mAbs incl. our CHOFlow® for afucosylated antibodies and the human GEX® platform for the development and production of complex glyco-biopharmaceuticals in the desired quality. A case study from a customer project demonstrates the suitability of our cell line platform for the production of complex bispecific mAbs in combination with bioprocess optimization capabilities.

Session Break to Transition into Plenary Keynote10:00

PLENARY KEYNOTE SESSION
基調講演(プレナリーセッション)

10:10

Introduction

Enkelejda Miho, PhD, Professor, Dean, University of Applied Sciences and Arts Northwestern Switzerland

10:15

Benchmarking the Impact of AI Biologics Discovery and Optimisation for Pharma

Rebecca Croasdale-Wood, PhD, Director, Augmented Biologics Discovery & Design, Biologics Engineering, Oncology, AstraZeneca

The biologics landscape is rapidly changing with the number of AI-enabled biologics in pre-clinical and clinical stages estimated to be 50-60 (1). This change is driven by the increase in enterprise software solutions to capture and store data, augmented discovery workflows, improvements in machine learning technology, and advances in computing power. Augmented biologics discovery has the potential to revolutionize biologics discovery, yet information of how in silico technologies perform, versus traditional discovery platforms is scarce. At PEGS Europe, we will present current in silico biologics design and optimisation technologies, with a focus on our internal efforts to benchmark the impact of combining novel in silico technologies with our existing biologics discovery platforms.

Coffee Break in the Exhibit Hall with Poster Viewing11:00

11:45

FEATURED PRESENTATION: Cell Line Development and Engineering Strategies for the Manufacture of Lentiviral Gene Therapy Viral Vectors

Ana Sofia Coroadinha, PhD, Lab Head, Health & Pharma Division, Animal Cell Technology Unit Cell Line Development and Molecular Biotechnology Lab, IBET

The gene and cell therapies market is growing, with several products being approved every year. The expression of viral vectors for gene therapy is  challenging however, since these often require the expression of toxic proteins posing obstacles in cell line development. This work discusses the main challenges lentiviral vector cell therapies face, and presents strategies and novel technologies to be adopted to enable their effective manufacture.

12:15

Baculovirus-Free Expression of Virus-Like-Particles in Insect Cells for Antibody Development

Maren Schubert, PhD, Research Group Leader, Department of Biotechnology, Technical University of Braunschweig

The baculovirus expression vector system leads to high yields of Virus-like-Particles (VLPs). Yet, it´s time-intensive, inflexible in regard of protein ratios, its quality can be hampered by low cell viability, and last but not least, purification is challenging due to simultaneously produced baculoviral particles. The here-presented alternative of baculovirus-free VLP production in insect cells avoids the pitfalls of BEVS and produces VLPs in high quality and quantity for antibody development.

12:45 Talk Title to be Announced

Speaker to be Announced

Session Break13:15

13:20 LUNCHEON PRESENTATION:GenScript, Not Only Gene Synthesis

Cristobal Almendros, PhD, Sales and Technical Leader - Spain and Portugal, GenScript Biotech (The Netherlands) B.V.

GenScript has earned its global reputation as the leading CRO in synthetic DNA. Yet, many customers remain unaware of our extensive range of services, including protein offerings. Leveraging state-of-the-art technologies and customization in collaboration with our customers, we provide an array of possibilities. Join us for an informative session where we showcase our comprehensive services, designed to support your recombinant expression protein projects and accelerate your scientific goals.

13:50 LUNCHEON PRESENTATION II:Strategies for Unveiling Optimal Bispecific Antibody Pairings

Julia Su, PhD, Associate Director - BD, WuXi Biologics, Protein Sciences, Protein Sciences, WuXi Biologics

Bispecific antibody production presents challenges like heterogeneity, production complexity, and stability issues, which can impact the quality of the final product. This presentation will disclose the innovative strategies to address these challenges in drug development. It includes the initial small-scale high-throughput production of a vast number of bispecific antibodies in identifying optimal pairings, as well as later stage large-scale production. Real-world case studies will showcase the successful application of these strategies.

Session Break14:20

OVERCOMING EXPRESSION AND PRODUCTION CHALLENGES FOR UNIQUE PROTEINS
固有のタンパク質における発現・生産の課題の克服

14:30

Chairperson's Remarks

Mercedes Marquez Martinez, PhD, Technical Coordinator & Acting Scientific Director, Protein Production Platform (PPP) - Nanbiosis, Autonomous, University of Barcelona (UAB)

14:35

KEYNOTE PRESENTATION: An Automated DNA Assembly Framework Enables Rapid and Scalable Plasmid Generation for Drug Discovery Applications

Robert G. Roth, PhD, Director, Protein Expression & Molecular Biology, Discovery Biology, R&D Biopharmaceuticals, AstraZeneca

Rapid and flexible construct generation at-scale is one of the most limiting first steps in the majority of drug discovery projects. The speed, quality, and cost of this process can be dramatically reduced by modular DNA design principles and automated fragmentation. To this end we have designed a robust, multi-module golden gate-based cloning platform for construct generation with a wide range of applications. In addition, to minimize timelines and cost for complex constructs, we developed a pipeline that performs fragmentation and codon optimization of long coding sequences in an automated manner.  

15:05

Host Comparative Production of Recombinant Proteins for Assembling as Nano- and Micro-Scale Materials for Drug Delivery

Mercedes Marquez Martinez, PhD, Technical Coordinator & Acting Scientific Director, Protein Production Platform (PPP) - Nanbiosis, Autonomous, University of Barcelona (UAB)

Recombinant proteins can be artificially self-assembled in the form of functional structures with promising applications in the field of drug delivery via nanostructured protein-only drugs. The antigenic RBD domain of the SARS-CoV-2 spike was produced in several expression systems such as bacteria, insect, and mammalian cells, to be used as a model to investigate the influence of the protein source in the production of nanoparticles and secretory microparticles. Both structures were generated in all cases but with different biophysical properties indicating thus that the selected expression system is a relevant factor for protein assembly into supramolecular structured and functional materials.

15:35 Talk Title to be Announced

Alexandra Baer, PhD, R&D Manager for Upstream Development of Mammalian Cells, Recombinant Proteins, Bioneer A/S

15:50 From Modular DNA Assembly to Recombinant Protein Production at Polyplus

Marine Houdou, Genetic Engineering Specialist, Polyplus

With recent acquisitions of e-Zyvec and Xpress Biologics, Polyplus offers now a unique integrated pDNA service with plasmid engineering and manufacturing. Our unique DNA assembly technology and proprietary software allow the tailor-made design and production of any plasmid. Synergistically, the 30+ years of combined expertise at Xpress Biologics reinforces our services at Polyplus to support recombinant protein manufacturing from 100 mg to 50 g at Research, GLP and GMP grade.

Refreshment Break in the Exhibit Hall with Poster Viewing16:05

17:00

Venom on Demand: Optimizing Snake Toxin Yield, Folding, and Purity in E. coli and P. pastoris with Biotinylation, Solubility, and Purification Tags

Esperanza Rivera de Torre, PhD, Assistant Professor, Center for Antibody Technologies, Department of Bioengineering, Technical University of Denmark

Animal venoms contain a plethora of biologically active toxins that have the potential to revolutionize antivenom development. However, producing functional toxins with the correct disulfide pattern is challenging. Our approach to engineering co-chaperon systems in Escherichia coli and leveraging Pichia pastoris' secretory capacity to produce natural and designed toxins. Our optimized strategies allow efficient toxin folding and purification, enabling downstream biotechnological applications, such as directed biotinylation for phage display.

17:30

Development of a Broadly Neutralizing Intranasal Anti-SARS-CoV2 Trimeric Sherpabody

Anna R. Makela, PhD, Senior Scientist, Department of Virology, University Of Helsinki

There is a need for prophylactic SARS-CoV-2 blocking agents that are invulnerable to mutational viral variation and economical to produce. TriSb92 is a highly manufacturable and stable trimeric antibody-mimetic sherpabody targeted against a conserved region of the viral spike glycoprotein. TriSb92 potently neutralizes SARS-CoV-2, including the latest Omicron subvariants. In mice, intranasal administration of TriSb92 as early as 8 hours before but also 4 hours after SARS-CoV-2 challenge can protect from infection. Triggering of a conformational shift in the spike trimer was revealed as the inhibitory mechanism. TriSb92 could be useful as a nasal spray for protecting from SARS-CoV-2 infection.

18:00

Recombinant Protein L: Production, Purification and Characterization of a Universal Binding Ligand

Oliver Spadiut, PhD, Associate Professor, Integrated Bioprocess Development, TU Wien, Vienna

Protein L (PpL) is a universal binding ligand that can be used for the detection and purification of antibodies and antibody fragments. Due to the unique interaction with immunoglobulin light chains, it differs from other affinity ligands, like protein A or G. However, due to its current higher market price, PpL is still scarce in applications. We investigated the recombinant production and purification of PpL and characterized the product in detail. In my talk I present a comprehensive roadmap for the production of the versatile protein PpL in E. coli.

18:30 PANEL DISCUSSION:

Protein Production Lab Challenges: Methodologies, Strategies, and the Art of Expressing Recombinant Proteins

PANEL MODERATOR:

Richard Altman, MS, Field Application Scientist, Life Science Solutions, Thermo Fisher Scientific

Protein expression laboratories provide crucial support to drug discovery efforts.  This panel discussion will focus on the concepts, technologies, and strategies necessary to meet the ever-increasing need for recombinant proteins.      

  • Know your protein!         
  • Strategies on how to manage multiple “top priority” projects        
  • Total workflow efficiency         
  • The importance of tech development to long term success       
  • Troubleshooting strategies or how much time should be spent before moving to the next option?
PANELISTS:

Nicola Burgess-Brown, PhD, Director of Enzymology and Protein Engineering, Exact Sciences Innovation

Bjorn Voldborg, MSc, Head, National Biologics Facility, DTU Bioengineering, Technical University of Denmark

Close of Optimising Expression Platforms Conference19:00


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