Protein Purification Technologies Track Banner


タンパク質精製技術
品質向上に向けたプロセスの合理化

このカンファレンスプログラムでは、タンパク質精製プロセスを合理化し、高まる需要に対応するため導入されている戦略や技術について考えます。

またスケールの問題、プロテインAやクロマトグラフィーなど既存技術の刷新、哺乳類細胞や細菌細胞、昆虫細胞など、さまざまな発現系でのタンパク質の精製についても議論します。

Final Agenda

11月16日 (木)

12:30 Registration

13:00 Dessert Break in the Exhibit Hall with Poster Viewing

抗体精製

13:30 Chairperson's Opening Remarks

Christopher H. Gray, Team Leader, Structural Biology, Drug Discovery Program, Cancer Research UK, The Beatson Institute

 

13:35 KEYNOTE PRESENTATION:
Protein Production Needs for Therapeutic Antibody Discovery and Development

Karin Felderer, Ph.D., Associate Director, Protein Production, Protein Sciences & CMC, MorphoSys AG

Protein production needs for therapeutic antibody projects are manifold. They span a wide range of scales from small scale high-throughput for early in vitro characterization up to gram scale productions for in vivo characterization and manufacturability assessment. In addition to antibodies and antibody fragments, various high quality antigens and tool proteins are required to facilitate successful projects. Case studies and strategies will be presented covering this spectrum of applications.

14:05 Exploration of an Atypical 'Platform' mAb and Resolution of Its Non-Ideal Behaviour during Downstream Process Development

Karolina Les, Ph.D., Scientist II, Purification Process Sciences, Biopharmaceutical Development, MedImmune, Ltd.

Efficient development of monoclonal antibodies (mAbs) is often founded on platform-based optimization of processes, allowing timely delivery of scalable and robust processes. While existing developability frameworks allow progression of best candidates into development and typically assure a good platform fit, cases of low platform compliance are unavoidable. The talk will discuss reasons behind an atypical behaviour of an antibody during purification and its mitigations. The main challenges included severe precipitation during purification and unusually high host cell protein (HCP) levels observed during early development.

Wacker Biotech14:35 Process Optimisation for Manufacturing of ScFv-Class Antibody with FOLDTEC®

Katrin Schweinitzer, Head, Downstream Processing Development, BioProcess Development, Wacker Biotech GmbH

Wacker Biotech will present a case study for development and optimization of a scalable process employing microbial system to produce the scFv-class antibody for commercial application. Using its E. coli-based FOLDTEC® technology, WACKER completely overhauled a process employed by customers to facilitate production on an industrial scale. WACKER's proprietary refolding technology was able to produce the desired product in enhanced yields and streamline the purification process with achieving higher purity than was hitherto possible.

15:05 Refreshment Break in the Exhibit Hall with Poster Viewing

抗体精製 (続き)

15:50 Streamlining Antibody Purification: A Novel Approach by Caprylic Acid-Based Impurity Precipitation

Stefan Schmidt, Ph.D., MBA, Senior Vice President, Process Science and Production, Rentschler Biotechnology

Benefits of implementing caprylic acid- (CA-) based precipitation in antibody purification are presented. Combining the common low pH viral inactivation step with CA precipitation greatly improves HCP clearance in different mAb processes. We show that CA precipitation can be applied as an excellent alternative to a polishing chromatography using case studies. The step constitutes a simple, robust and economic step in a mAb purification process resulting in increased purification performance and higher safety of two-column processes.

16:20 A Microfluidic Toolbox for Screening and Downstream Process Optimization

M. Raquel Aires-Barros, Ph.D., Professor, Institute for Bioengineering and Biosciences (IBB), Bioengineering, Instituto Superior Tecnico, Universidade de Lisboa

Antibodies and other protein products are biopharmaceuticals of critical importance that need to be purified in a cost effective and efficient manner. Here, a methodology for the rapid screening of antibody extraction conditions using a microfluidic channel-based toolbox is presented which can also be used for process development. A first microfluidic structure allows a simple negative-pressure driven rapid screening of up to 8 extraction conditions simultaneously, while a second microfluidic structure allows the integration of multi-step extraction steps.

16:50 End of Day

17:00 Dinner Short Course Registration

17:30-20:30 Recommended Short Course*

SC7: Protein Purification Strategies

*Separate registration required

11月17日 (金)

08:00 Registration and Morning Coffee

膜タンパク質の精製

08:30 Chairperson's Remarks

Stefan Schmidt, Ph.D., MBA, Senior Vice President, Process Science and Production, Rentschler Biotechnology

08:35 Do We Need to Purify Membrane Proteins? Expression Tricks and Solid-State NMR Spectroscopy in Native Membranes

Dirk Linke, Ph.D., Professor, Molecular Microbiology, Biosciences, University of Oslo

In recent work, we have developed E. coli expression strains that lack multiple of the most abundant outer membrane proteins. This enables us to perform solid-state NMR spectroscopy directly on native membranes after overexpression of targets in 13C/15N labeling medium. Membrane protein purification becomes obsolete in this procedure, and the proteins can be studied in their native environment without risk of denaturation.

09:05 The Salipro Lipid Nanoparticle System for Detergent-Free Stabilization of Membrane Proteins

Jens Frauenfeld, Ph.D., 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. Salipro Biotech has developed a novel system to reconstitute membrane proteins into self-assembling lipid nanoparticles. The Salipro system is applicable for various classes of membrane proteins (channels, receptors, transporters, etc). Salipro nanoparticles stabilize membrane proteins in a lipid environment and allow working in detergent-free buffer systems.

09:35 Membrane Proteins: From Lab-Scale towards Large-Scale Production

Julie Bomholt, Ph.D., Head, Downstream Processing and Quality Control, Downstream Processing, Golgi ApS

By use of the physiologically important trans-membrane protein family of aquaporins as model proteins, we developed a S. cerevisiae based expression system, and identified conditions that allowed us to substantially increase the membrane density of recombinant functional aquaporin. We further developed a purification scheme that allowed us to isolate high-yield preparations of pure and functional aquaporin. Based on this, we conducted a comparative study of human aquaporins from expression level to functional reconstitution of purified detergent stabilized protein.

10:05 Coffee Break with Poster Viewing

精製にまつわる課題の克服

10:35 Production of Human Aldehyde Oxidase (AO) in Baculovirus-Infected Insect Cells (BEVS)

Ciaran N. Cronin, Ph.D., Associate Research Fellow, Head, Parallel Protein Production Group, and Group Leader, Gene-to-Structure, Pfizer Global R&D

Aldehyde oxidase (AO) is a key enzyme activity to consider during the development of small molecule human therapeutics. There are numerous literature examples of failed clinical drug trials due to a lack of preclinical testing of potential drug exposure to AO activity. This presentation summarizes the numerous design, expression and purification experiments that were undertaken in order to optimize the production of recombinant human AO for deployment in Pfizer's screening cascade.

11:05 Human Heparanase: Lessons Learned from a Self-Destructing Project

Mario Lebendiker, Ph.D., Head, Protein Purification Facility, Wolfson Center for Applied Structural Biology, Hebrew University of Jerusalem

Our case study describes the challenges encountered in each step of crystallizing Human Heparanase (hHep), an endo-beta-D-glucuronidase. We needed to overcome irreproducible expression productivity of this "self-destructing" protein in insect cells, followed by optimizing a purification strategy for this non-tagged target with conventional chromatography. We then faced batch-to-batch variability in crystallization, and finally, an inability to solve collected data from crystals, until we produced a fusion protein construct with known structure to obtain phases.

11:35 High-Throughput Knockout of Difficult-to-Remove and Troublesome CHO Host Cell Proteins to Create a Clean CHO Cell Line

Stefan Kol, Ph.D., Protein Biochemist, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark

The CHO Cell Line Engineering department is addressing the need to obtain high yields and quality of biopharmaceuticals produced in optimized CHO cells through genome-scale-based methodologies. Although many host cell protein (HCP) impurities are effectively removed in downstream purification processes, a small population of HCPs are particularly challenging. Here, we present our efforts to identify and knock out several CHO HCPs and our characterization of the resulting cell line.

12:05 Sponsored Presentation (Opportunity Available)

12:35 Problem-Solving Breakout Discussions with a Light Snack in the Foyer

13:35 Session Break

新たな技術

14:00 Chairperson's Remarks

Jens Frauenfeld, Ph.D., CEO, Salipro Biotech AB

14:05 Novel Single-Column Simulated Moving-Bed Chromatography for Quasi-Continuous Purification of Biomolecules

Jose Paulo Mota, Ph.D., Professor, Chemical and Biochemical Engineering, LAQV-REQUIMTE, Chemistry, Science and Technology, Universidade NOVA de Lisboa

14:35 Parallel Rapid Expression & Purification of Proteins for Crystallography (PREPX)

Michael Fairhead, Ph.D., PDRA, Structural Genomics Consortium, University of Oxford

Pipelines for high-throughput cloning and small-scale expression and purification are well established. However, subsequent scale-up and purification often involves only working with a few pipeline hits (usually the easiest to produce) and these are not always suitable for crystallography. PREPX is a medium-throughput approach to evaluate larger numbers of clones (48 per week), and their suitability for producing diffraction quality crystals.

15:05 Expressed Protein Ligation: A New Paradigm as a Reagent Platform for Preclinical Drug Discovery

Rosalie Matico, Associate Fellow, Investigator, Protein Cellular and Structural Sciences, GlaxoSmithKline plc

Expressed protein ligation (EPL) utilizes genetically engineered inteins to join two protein (peptide) fragments via a native peptide bond. Ligation occurs through an intein mediated C-terminal thioester on one protein fragment and an N-terminal Cys-SH on a second protein or peptide fragment. We developed a reagent platform by harnessing (EPL) to specifically label proteins at the C-terminus with a cysteine-lysine dipeptide chemically modified at the lysine with biotin, fluorescein, peglyation, etc., prior to ligation.

精製プロセスの改善

15:35 Optimization of Secreted Target Protein Yields by Introduction of in silico Designed Stabilizing Mutations

Svend Kjaer, Ph.D., Deputy Head, Structural Biology Science Technology Platform, Francis Crick Institute

Protein stability is a fundamental and crucial parameter in a range of applications from crystallization to biotherapeutics. We present data of how a few stabilizing mutations, designed by the PROSS algorithm based on crystal structures and multiple sequence alignments, increase the Tm of a misfolding-sensitive secreted protein by 20°C. We present data showing how protein expression in higher eukaryotic systems is significantly improved by the stabilization.

16:05 Tandem-Tag Vectors for Enhanced Soluble Yield, Expression Monitoring and Streamlined Purification

Christopher H. Gray, Team Leader, Structural Biology, Drug Discovery Program, Cancer Research UK, The Beatson Institute

Our suite of novel, tandem-tagged Escherichia coli vectors offers multiple options for increased soluble expression, and highly efficient purification. Dual promoters allow the simultaneous expression of an MBP-target fusion and a tag cleaving protease. Similarly, this in vivo cleavage system functions with GFP-target fusions allowing fluorescent tracking of recombinant expression during culture. Finally, we have designed a set of tandem-affinity tagged vectors generating protein that can be highly purified in a single step.

16:35 End of Conference

* 不測の事態により、事前の予告なしにプログラムが変更される場合があります。

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