Cambridge Healthtech Institute’s Tenth Annual

Protein Purification and Recovery

Streamlining & Innovating Processes

January 9-10, 2018

 

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

Streamlining Protein 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 Introduction of a Disruptive Technology into a Highly Regulated Industry: A Brief History with Lessons Learned

David WoodDavid Wood, Ph.D., Professor, Chemical & Biomolecular Engineering, The Ohio State University

Over the past 20 years, we and others have worked to develop a highly disruptive self-cleaving tag technology for the biopharmaceutical industry. As we draw closer to this goal, it is worthwhile to consider the larger picture of how we have adapted our goals and approaches to this highly regulated and rapidly developing industry. This talk will cover the history and most recent developments of this technology in this context.

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

Sophia_HoberSophia 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.

3:15 SELECTED POSTER PRESENTATION:
Designing a State-of-the-Art Sample Management Facility to Empower Research

Katharine_HeeringaKatharine Heeringa, Scientific Researcher, Protein Sciences, Genentech, Inc.

Biomolecular Resources Sample Management (BMR-SM) at gRED is a state-of-the-art facility supporting all of research at Genentech, acting as both a nucleic acid and protein repository and a nucleic acid production facility. BMR-SM leverages bioinformatics and automation to support multiple users and at multiple access points with a diverse range of products and comprehensive sample and process tracking. We seek to build a core biologic molecule repository and nucleic acid production group that can expand and adapt to an ever-changing research environment.

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

PURIFYING ANTIBODIES

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_ZhangQingchun Zhang, Ph.D., Senior Scientist, Process Development, Amgen, Inc.

Recent advances in mass spectrometry allow the identification and quantification of individual HCPs. These advancements now make it possible to characterize HCPs in the presence of mAbs and to collect data to perform a risk assessment of individual HCP impurities. A comprehensive comparison of HCP subpopulations across different mAbs was conducted and the potential immunogenicity risk of commonly observed HCPs was investigated using both in silico prediction and an in vitro PBMC-based assay.

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

Greg_WeissGregory 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_GrayChristopher 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.

 

Nanotemper Technologies 9:35A Quick Check of Protein Quality that Will Vastly Improve all Protein Purification and Characterization Workflows

Fung PeterPeter Fung, Ph.D., Senior Manager, Product Marketing, NanoTemper Technologies

Starting with material of questionable quality for protein purification and characterization leads to irreproducible or ambiguous results. Methods such as chromatography while widely used, can also derail experiments—due to the amount of time and expertise required to perform these techniques. We present a new platform that swiftly identifies sample quality and relative functionality in minutes complementing and guiding purification and characterization workflows—making go/no go decisions easy and quick—saving time, effort and cost downstream.

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

NEXT-GEN CHROMATOGRAPHY

10:50 SELECTED POSTER PRESENTATION:
Tricks to Enhance Bispecific Antibody Purification by Using Protein L Chromatography

Yuichiro_ShimizuYuichiro Shimizu, Ph.D., Research Manager, Chugai Pharmabody Research PTE. LTD.

In this study, we have established a system to efficiently separate 4-chain bispecific antibodies (BiAb) as well as to remove certain type of by-products by using protein L chromatography. Due to its simplicity and robustness, this method can potentially be used broadly for BiAb purification with minimum engineering of amino acid sequences.

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

Dmitry_VassylyevDmitry 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.

iba  12:20pm 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.

12:50 LEnjoy 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_RothnieAlice 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_YeliseevAlexei 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

Soeren_Flygenring_BassetSøren 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_DumetzAndré 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_BantaScott 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


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