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The booming field of biologics places ever greater demands on protein production. Whether for research or to create biotherapeutics, scientists must purify protein following expression and scale-up.  To streamline processes, researchers need to integrate upstream and downstream phases, and reduce the number of steps required to achieve pure, properly-folded protein. A plethora of technologies and tools are available to support purification, but which are the best? As protein-based molecules become more complicated, they pose greater challenges for purification. Bispecifics, ADCs, fusion and membrane proteins present purification obstacles that may not have been tackled before. And aggregation continues to be a nagging issue waiting to trip up an earnest researcher. This conference presents the tricks and technologies used by purification leaders who have refined techniques that remove headaches from protein processing. Experts will address ‘traditional’ technologies as well as new approaches, and will touch on issues of scale, as well as purifying protein in various expression systems; all with an eye to streamline purification while ensuring quality.


Final Agenda

THURSDAY 15 NOVEMBER

13:00 Registration

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


INNOVATING PURIFICATION PROCESSES

14:00 Chairperson’s Opening Remarks

Dorota Antos, PhD, Professor, Chemical and Process Engineering, Rzeszow University of Technology


14:05 KEYNOTE PRESENTATION: How Problems of Protein Purification Are Being Addressed across Structural Laboratories in Europe: Insights from the European Research Infrastructure Consortium

Owens_RayRay Owens, PhD, Professor, Molecular Biology and Head, Oxford Protein Production Facility, University of Oxford

Technology developments to streamline the production of proteins for structural biology have been largely driven by the demands of structural proteomics. However, the purification of increasingly complex proteins and protein assemblies has challenged traditional high-throughput structural proteomic workflows. The response of a number of academic centres in Instruct, a distributed European Research Infrastructure Consortium (www.structuralbiology.eu) will be reviewed and common trends highlighted.

14:35 Simplification of Purification Strategies for Mammalian Proteins from Single Targets to High-Throughput Projects Using an Enhanced Biochemical Code

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

We are investigating the sequences of amino acids and their associated post-translational modifications that confer superior transport characteristics upon secreted proteins to move through and across gradients. With one eye on creating new protein engineering design principles, we are aiming to understand the elements of encoded behaviour of proteins including IgG, cytokines, interferons and the very many proteins that make up our secretome using a high-throughput expression and interrogation model in HEK293. Establishing high-throughput expression, purification and transport assays will be described.

15:05 Overcoming Limitations of Conventional Tag Systems – Strep-Tactin®XT Applications

Dennis Karthaus, MSc, IBA Lifesciences

The Strep-Tactin®XT:Twin-Strep-tag®-purification system enables protein purification at high yields and purity under physiological conditions. Providing the highest binding affinity among all affinity tag systems, the technology fulfills the demands of mammalian expression systems (e.g. Expi) and is well suited for downstream applications like SPR.

15:20 Sponsored Presentation (Opportunity Available)

15:35 Networking Refreshment Break


HARNESSING THE POWER OF SMALL

16:00 Purification of Fabs from E. coli Cell Lysates – A Tricky Endeavour

Spadiut_OliverOliver Spadiut, PhD, Assistant Professor, Chemical, Environmental and Biological Engineering, Integrated Bioprocess Development, Technische Universität Wien (TU Wien)

In my presentation, I will highlight the challenges when purifying added value molecules, like Fabs, from E. coli cell lysates and compare different strategies to do so. Our group’s new feeding approach allows expression of complex products as soluble and active protein. Cell viability and growth can be prolonged by this approach which leads to higher overall yields and thus lower production costs. Our platform technology accelerates bioprocess development and yields higher product titers in E. coli.

16:30 Protein Purification and Detection with Nanobodies

Rothbauer_UlrichUlrich Rothbauer, PhD, Professor, Pharmaceutical Biotechnology, Natural and Medical Sciences Institute, University of Tübingen

Nanobodies are attractive tools for protein purification, detection and analysis as they are small, highly stable, and are easily producible thereby offering an unlimited supply of consistent binding molecules. Recent advances in identification of target-specific nanobodies from synthetic gene libraries makes these tools broadly available. Here, we present our latest progress in nanobody generation, functionalization and application for protein purification and detection.

17:00 End of Day

17:30 Dinner Short Course Registration*

17:3020:30 Dinner Short Courses


Recommended Short Course*

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

O'Connell_DavidDavid 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.

FRIDAY 16 NOVEMBER

08:00 Registration and Morning Coffee


PURIFYING MEMBRANE PROTEINS

08:30 Chairperson’s Remarks

Maximilian Hartl, PhD, Senior Scientist, Roche Pharma Research & Early Development (pRED), Large Molecule Research, Roche Innovation Center Munich

08:35 Structural Biology of Membrane Proteins: Expression Tricks, and to Solubilize or Not to Solubilize

Linke_DirkDirk Linke, PhD, Professor, Genetics and Evolutionary Biology, Biosciences, University of Oslo

In recent work, we have designed expression systems for bacterial outer membrane and surface proteins. These can be used to express and purify, e.g., important vaccine candidates for bacterial diseases, but also for studying membrane protein structure directly in the membrane with solid-state NMR methods.

09:05 Adaptive Automated Membrane Protein Purification Using AkTA Avant

Lee_JonasJonas Lee, PhD, Scientist, Protein Technologies, Amgen, Inc.

Transmembrane proteins are key targets in drug discovery. However, they are difficult to purify due to complex buffer requirements to solubilize. We use various high-throughput methods to screen for best detergent conditions followed by innovative methods to purify multiple targets in different buffer conditions automatically.

09:35 Nanoparticles to Stabilize Membrane Proteins in a Lipid Environment

Frauenfeld_JensJens Frauenfeld, PhD, CEO, Salipro Biotech AB

More than 60% of all current drugs target membrane proteins. However, membrane proteins are very unstable, which is a major challenge for the pharmaceutical industry. Here we present the latest results on a novel system to stabilize membrane proteins using Salipro nanoparticles. Salipro nanoparticles stabilize membrane proteins in a lipid environment that allows them to work in detergent-free buffer systems.

10:05 Networking Coffee Break


NEXT-GEN CHROMATOGRAPHY

10:35 Potential of Centrifugal Partition Chromatography for the Separation of Proteins

Minceva_MirjanaMirjana Minceva, PhD, Assistant Professor, Biothermodynamics, Life and Food Sciences Weihenstephan, Technische Universität München

Centrifugal partition chromatography (CPC) is a solid support-free chromatography method in which both stationary and mobile phase are liquid. This technology combines advantages of liquid-liquid extraction and chromatography, such as high loading capacity, high recovery and high resolution. Moreover, as a result of the liquid nature of the stationary phase, problems associated with column packing are avoided. CPC can be used with aqueous two-phase systems (ATPS) providing a mild environment for the separation of proteins. In this talk, the potential of CPC will be demonstrated for the separation of model mixture of proteins.

11:05 Nanofibre Based Protein A Chromatography for a Truly Disposable Unit Operation

Bracewell_DanDaniel Bracewell, PhD, Professor, Bioprocess Analysis, Biochemical Engineering, University College London

A cellulose nanofibre adsorbent has been designed with dynamic binding capacities independent of residence time, thus enabling a rapid cycling chromatography mode of operation, with each unit being cycled >100 times in a single working shift. The case study presented covers early stage process development of a mAb purification process from an industrial clarified feedstream. The work demonstrates feasibility and scalability for industrially relevant single-use product capture utilising the powerful protein A ligand at a much greater throughput and therefore productivity.

11:35 Sponsored Presentation (Opportunity Available)

12:05 Problem-Solving Breakout Discussions with a Light Snack


IMPROVING AND SCALING UP PURIFICATION PROCESSES

13:00 Chairperson’s Remarks

Ulrich Rothbauer, PhD, Professor, Pharmaceutical Biotechnology, Natural and Medical Sciences Institute, University of Tübingen

13:05 Process Development of the Antibody-Drug Conjugate (ADC) SYD985 – A Case Study

Guy De Roo, PhD, Lead Scientist, Synthon Biopharmaceuticals BV

SYD985 is an antibody-drug conjugate (ADC) based on trastuzumab and a cleavable linker-duocarmycin payload. A case study will be presented in which a hydrophobic interaction chromatography (HIC) purification process was developed allowing removal of the undesired antibody species together with unbound linker-drug. It was possible to elute the product (SYD985) using mild conditions without requirement for any organic solvent. The HIC purification step was scaled up demonstrating consistency and robustness.

13:35 New Formats, New Experiences - DSP Feedback for Carefully Selecting a Molecule for Development

Hartl_MaximilianMaximilian Hartl, PhD, Senior Scientist, Roche Pharma Research & Early Development (pRED), Large Molecule Research, Roche Innovation Center Munich

Biopharmaceuticals evolved from copying natural molecules to tailor-made, highly engineered drugs with disease specific action modes. The impressive ideas of our molecule designers often result in promising early in vivo data that challenge technical-scale drug development. In this talk, we show examples of non-predicted challenges we faced during purification of novel drugs. We present solutions and feedback from purification for the selection process of next-generation drugs.

14:05 The Effect of New Stabilizers in Downstream mAb Process Intermediates

Irina Ramos, PhD, Downstream Process Scientist, MedImmune, Inc.

14:35 Pitfalls in Design and Scaling Up Protein Chromatography

Antos_DorotaDorota Antos, PhD, Professor, Chemical and Process Engineering, Rzeszow University of Technology

Because of the complexity of thermodynamic, kinetic, and hydrodynamic effects accompanying protein chromatography, design and scaling up of the process is often impossible without understanding underlying adsorption mechanism. Specific effects that can occur in protein chromatography will be discussed. The phenomena of band deformation due to mass transport resistances, protein unfolding, sample solvent effect, and dispersion in extra column volumes will be illustrated. The solvent gradient and temperature mediated separations will be considered.

15:05 Monoclonal Antibody Reduction and Re-Oxidation by Copper Sulfate during Manufacturing and Impact on Product Quality

Nilapwar_SanjaySanjay Nilapwar, PhD, Senior Research Investigator, Large Molecule Purification Development, Incyte Corporation

Significant disulfide bond reduction of an IgG1 monoclonal antibody was observed during the late stage of the CHO cell culture in manufacturing, leading to the batch failure due to significant amount of low molecular weight species and aggregates. Two methods of copper sulfate spiking were investigated to prevent and reverse the product disulfide bond reduction during manufacturing. Both methods could re-oxidize the reduced product and prevent further reduction throughout the manufacturing process.

15:35 End of Summit

* 活动内容有可能不事先告知作更动及调整。

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