Cambridge Healthtech Institute's Tenth Annual

Protein Purification and Recovery

( 蛋白质的纯化与回收 )

Streamlining & Innovating Processes

2018年1月9日 - 10日

 

Cambridge Healthtech Institute's Protein Purification and Recovery conference examines the strategies that efficiently lead to pure protein for research or therapeutic use. As the most costly and time-consuming process in the manufacturing of protein-based therapies, purification poses continual challenges for streamlining steps and cutting costs. Challenges are multiplied when purifying complex molecules, such as membrane proteins, bispecifics and antibody-drug conjugates. This leading purification meeting explores how experts are optimizing processes to reach project goals in a timely way. Along with innovating 'traditional' technologies such as affinity tags, Protein A, and chromatography, leaders will also address alternatives and breakthroughs, such as continuous processing.

Final Agenda

 

TUESDAY, JANUARY 9

1:00 pm Registration

1:30 Refreshment Break in the Exhibit Hall with Poster Viewing

Antibody Purification

2:00 Chairperson's Opening Remarks

Christopher H. Gray, Ph.D., Team Leader, Structural Biology, Drug Discovery Program, CRUK Beatson Institute

KEYNOTE PRESENTATION

2:05 Rapid High-Resolution Separation and Analysis of Monoclonal Antibody Samples Using Laterally-Fed Membrane Chromatography

Raja Ghosh, Ph.D., Professor and Canada Research Chair, Bioseparations Engineering, Chemical Engineering, McMaster University

Laterally-fed membrane chromatography (LFMC) devices combine high speed with high resolution in separation. These devices have low variability in flow-path lengths and consequently peaks obtained are very sharp, and the resolution of multi-component protein separation is comparable or sometimes even better than resin-based columns. The use of LFMC devices for high-resolution purification applications such as separation and analysis of monoclonal antibody aggregates and charge variants will be discussed.

2:45 Optimization of an IgG-Binding, Protein A-Based Purification Matrix

Sophia Hober, Ph.D., Professor, Molecular Biotechnology, KTH Royal Institute of Technology

Presented here is an engineered protein based on the Protein A-derived Z domain, to which a calcium-binding EF-loop has been introduced. The new protein domain, ZCa, is shown to have a calcium dependent binding to IgG and can be used to purify antibodies with elution by EDTA at pH 5.5, providing a very valuable new tool for antibody and Fc-fusion protein purification. The process of the development, its function as well as the molecular explanation of its behavior will be discussed.

iba 3:15 The Strep-tag® Technology - The Superior Tag System for the Entire Protein Production Workflow

Dennis_NiermeierDennis Niermeier, MSc, IBA Lifesciences

IBA is focused on a comprehensive product portfolio around its proprietary Strep-tag® technology that covers the entire protein production workflow from cloning, expression and purification as well as protein immobilization for assays. Especially our Strep-Tactin®XT is superior to other systems due to its extreme high affinity while maintaining reversibility.

3:45 Refreshment Break in the Exhibit Hall with Poster Viewing

4:30 An Introduction to HisMAB: An Antibody-Based Affinity Purification System for His Tagged Proteins

Jiansheng Wu, Ph.D., Principle Scientific Manager, Protein Chemistry, Genentech, Inc.

Ni based methods have been well established for the purification of his tagged proteins for decades. They usually have high binding capacity at relatively low cost. However, due to the low selectivity of Ni resin, sometimes it is difficult to purify His tagged proteins with poor expression. In my lab, we have developed a new antibody based affinity system for the purification of His tagged proteins. The antibody called HisMAB binds to his tagged proteins at high affinity. It has very high specificity toward his tagged proteins. In this talk, we will share our experience using the antibody. HisMAB is best suited for the purification of secreted his tagged proteins expressed by BEVS and mammalian systems.

5:00 Co-Elution of Host Cell Proteins with Monoclonal Antibodies and Their Potential Immunogenicity Risk Assessment

Qingchun Zhang, Ph.D., Senior Scientist, Amgen, Inc.

5:30 Close of Day

5:30 - 5:45 Short Course Registration

5:45 - 8:45 Dinner Short Courses*

* Separate registration required

WEDNESDAY, JANUARY 10

8:00 am Registration and Morning Coffee

innovating processes

8:30 Chairperson's Remarks

Sophia Hober, Ph.D., Professor, Molecular Biotechnology, KTH Royal Institute of Technology

FEATURED PRESENTATION

8:35 Ten-Minute Purification and Rapid Folding of Proteins by Vortex Fluidic Device

Gregory A. Weiss, Ph.D., Professor, Chemistry and Molecular Biology & Biochemistry, University of California, Irvine

Recombinant proteins often require process intensive purification and refolding steps. In collaboration with Professor Colin Raston (U. Flinders, Australia), my lab applies a vortex fluidic device for continuous flow purification and refolding of proteins. Cell lysates can be directly processed without chromatography or centrifugation steps. Furthermore, the proteins remain bioconjugated to the surface of the flow reactor, allowing in-line bioprocessing by enzymes after their recovery from lysates.

9:05 Bigger, Brighter, Faster: Accelerating Protein Production by Enhancing Soluble Yield, Monitoring Expression and Compressing Chromatography Strategies

Christopher H. Gray, Ph.D., Team Leader, Structural Biology, Drug Discovery Program, CRUK Beatson Institute

We increased output using auto-cleaving MBP fusions, elevating soluble expression while eliminating MBP from purification. Additionally, we developed systems for rapid monitoring of target expression during fermentation using a co-expressed GFP tracer. Finally, we developed multimodal style affinity chromatography for tandem tagged proteins giving high purity material in a single column without need for slow polishing steps. The net result is reliability, higher yields and accelerated delivery to downstream users.

9:35 Sponsored Presentation (Opportunity Available)

10:05 Coffee Break in the Exhibit Hall with Poster Viewing

NEXT-GEN CHROMATOGRAPHY

10:50 Liquid-Liquid Phase Separation Causes High Turbidity and Pressure during Low Ph Elution Process in Protein A Chromatography

Haibin Luo, Ph.D., Scientist II, MedImmune LLC

11:20 An Efficient, Ultra-High Affinity Chromatography in a One-Step Purification of Complex Proteins

Dmitry G. Vassylyev, Ph.D., Professor, Biochemistry and Molecular Genetics, University of Alabama at Birmingham

Protein purification is the basis for numerous biochemical and biomedical studies. It is particularly crucial and challenging for structural analysis and industrial protein production, where it has to meet the High-yield/High-purity/High-activity (HHH) requirement. The ultra-high affinity (CL7/Im7) purification system allows for one-step HHH-purification of a wide range of traditionally challenging proteins and might emerge as a universal high-throughput purification tool to advance biological studies and manufacturing of therapeutic proteins.

11:50 Multidimensional Chromatography Coupled with Mass Spectrometry Characterization of Species Observed in Native Separations of a Thiol-Linked Antibody-Drug Conjugate

Andrew Holloway, Senior Research Associate, Analytical Sciences, Seattle Genetics, Inc.

12:20 pm Sponsored Presentation (Opportunity Available)

12:50 Session Break

1:00 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

Purifying Membrane Proteins

2:00 Chairperson's Remarks

Dmitry G. Vassylyev, Ph.D., Professor, Biochemistry and Molecular Genetics, University of Alabama at Birmingham

2:05 Detergent-Free Purification of Membrane Proteins Using SMA Polymer

Alice Rothnie, D.Phil., Lecturer, Biochemistry, Life & Health Sciences, Aston University

Purification of membrane proteins can be challenging due to the need to remove them from the membrane. Traditionally, this is achieved using detergents, which often cause instability and/or loss of function. A new methodology for the extraction and purification of membrane proteins uses a styrene maleic acid co-polymer (SMA) which inserts in the membrane and assembles into small discs of bilayer encircled by polymer, termed SMA lipid particles (SMALPs). These particles are stable, maintain the lipid environment of a protein and are amenable to structural and biophysical studies.

2:35 Small Affinity Tags for Efficient Purification and Recovery of Integral Membrane Receptors

Alexei Yeliseev, Ph.D., Staff Scientist, LMBB, NIH/ NIAAA

We expressed the recombinant cannabinoid receptor CB2 expressed in E. coli cells as well as in expi CHO cells in milligram quantities. Protein was purified by tandem affinity chromatography using either His tag/twin Strep-tag or His tag/EF1 tag pairs. In this work, we compare the use of single affinity and tandem affinity purification strategies. The protocols developed in our laboratory can be applied to expression and purification of other membrane receptors for structural and functional studies.

3:05 Refreshment Break in the Exhibit Hall with Poster Viewing

OVERCOMING PERSISTENT CHALLENGES

4:00 The Development of Optimised Silica Resins to Solve Complex Purification Challenges

Soren Flygenring Basset, Ph.D., Director, R&D, Novo Nordisk Pharmatech A/S

4:30 Establishing Guiding Principles to Optimize Host Cell Protein Removal during Purification Process Development

Andre C. Dumetz, Ph.D., Senior Scientific Investigator, Biopharm Downstream PD-3, R&D Platform Technology & Science, GlaxoSmithKline

5:00 Engineering the Beta Roll Domain for Bioseparations Applications

Scott Banta, Ph.D., Professor, Chemical Engineering, Columbia University

RTX peptide domains are intrinsically disordered and reversibly fold into the beta roll secondary structure domain specifically upon calcium addition. RTX domains created from concatenated consensus sequences reversibly precipitate with calcium addition and we have developed this for non-chromatographic protein purification. We have also engineered a face of the beta roll domain to bind to a target protein and this enables a new calcium-dependent catch and release affinity chromatography platform.

5:30 - 6:45 Networking Reception in the Exhibit Hall with Poster Viewing

6:45 Close of Protein Purification and Recovery Conference

* 活动内容有可能不事先告知作更动及调整。