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Cambridge Healthtech Institute’s 11th Annual

Protein Purification and Recovery

Streamlining and Innovating Processes

January 15-16, 2019

 

 

In the world of biologics, purifying protein remains a constant bottleneck and nagging headache. A process that works great for one protein, may not work at all for the next. Not only are the tasks challenging, but outcomes must be ensured to result in properly folded protein. CHI’s Protein Purification and Recovery conference examines the strategies that efficiently lead to pure protein for research or therapeutic use. This leading conference illustrates how ‘traditional’ strategies (protein A, chromatography, affinity tags) are being innovated and improved, while also demonstrating the new technologies that are being introduced and integrated to help streamline purification while ensuring quality. This conference will also explore the finesse required when purifying complex molecules, such as membrane proteins, bispecifics and antibody-drug conjugates, in the ever-present quest for purity.

Final Agenda

 

TUESDAY, JANUARY 15

1:00 pm Registration (Sapphire West Foyer)

1:30 Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

Purifying Antibodies
Aqua EF

2:00 Chairperson’s Opening Remarks

Peter Schmidt, PhD, Director, Recombinant Technologies Research, CSL Behring


KEYNOTE PRESENTATION

2:05 Evaluation of Recent Innovations for Capture of Antibodies and Antibody-Like Biotherapeutics

Alan K. Hunter, PhD, Director, Purification Process Sciences, MedImmune, LLC

As therapeutic use of monoclonal antibodies and related molecules continues to grow, affinity chromatography remains the primary capture modality due to high specificity, platformability, and strong regulatory track record. In this work, we provide a comprehensive evaluation of next-generation Protein A stationary phases for biotherapeutic manufacture. Lastly, we discuss purification strategies for bispecific antibodies with a mAb-like architecture using light chain affinity chromatography.

2:45 Replacing Protein A/G with Nucleotide Binding Site Ligands on Resins and Membranes for Chromatography and Spin Columns for Antibody Purification

Bilgicer_BasarBasar Bilgicer, PhD, Associate Professor, Chemical and Biomolecular Engineering, Mike and Josie Harper Cancer Research Institute, NDnano Center for Nano Science and Technology, University of Notre Dame

The traditional techniques of protein A/G affinity chromatography for antibody purification have well established limitations commonly overlooked due to convenience and absence of reliable options. We utilize the conserved nucleotide-binding site (NBS) of immunoglobulins to enable capturing of antibody on an affinity column. The results reveal >99% column efficiency with >99% purity for antibodies, suggesting that the NBS column is a universal, stable, reusable, and inexpensive alternative for purification of humanized and chimeric antibodies.

GE_Healthcare 3:15 Driving Efficiency in Purification
with Automated Multistep and Parallel Chromatography

Hoang Tran, Senior Field Applications Scientist, GE Healthcare Life Sciences

The influence of process intensification in research is driving scientists to improve upon the traditional single-step purification methodology, moving them into more efficient and automated parallel and continuous processing. Implementation of automated multistep and parallel purification processes have increased workflow productivity and efficiency leading to lowering cost. This presentation will focus on case studies of these advanced chromatography automation techniques, showing the promise of process intensification in the research environment.

 

3:45 Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

Purifying Antibodies (Cont.)
Aqua EF

4:30 Purification of Common Light Chain IgG-Like Bispecific Antibodies Using Highly Linear pH Gradients

Beth Sharkey, Scientist I, High Throughput Expression, Adimab, LLC

A variety of bispecific constructs benefit from the use of a single variable light region pairing with multiple distinct variable heavy regions. This talk will demonstrate new techniques to purify these common light chain bispecific IgG molecules to homogeneity. Data will be shared on the production of a panel of bispecific antibodies that bind each target with high affinity and exhibit favorable biophysical properties, similar to traditional therapeutic antibodies.

5:00  Development and Optimization of Challenging Unit Operations with Line of Sight to Manufacturing for a Shear Sensitive, Aggregation Prone, & Low pI Monoclonal Antibody

Rios_SandraSandra E. Rios, PhD, Principal Scientist, Downstream Process Development and Engineering, Merck & Co.

Hydrophobic interaction chromatography (HIC) is commonly used as a polishing step in monoclonal antibody purification processes. HIC offers an orthogonal selectivity to ion exchange chromatography and can be an effective step for the clearance of aggregate and other process-related impurities. This study focused on the development and optimization of challenging unit operations with line of sight to manufacturing for a monoclonal antibody with the unique characteristics of low pI, self-association prone and shear sensitivity.

5:30 Close of Day

5:30 - 5:45 Short Course Registration (Sapphire Ballroom)


5:45 - 8:45 Recommended Dinner Short Courses*

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*Separate registration required

 

WEDNESDAY, JANUARY 16

7:45 am Registration and Morning Coffee (Sapphire West Foyer)

Continuous Technology for Antibody Purification
Aqua EF

8:15 Chairperson’s Remarks

Yuyi Shen, PhD, Principal Scientist, Technical Department, Grifols, S.A.

FEATURED PRESENTATION

8:20 Continuous Downstream Processing of Monoclonal Antibodies

Zydney_AndrewAndrew Zydney, PhD, Distinguished Professor, Chemical Engineering, The Pennsylvania State University

There is growing interest in the development of integrated continuous bioprocesses with enhanced productivity and greater flexibility. This talk will present several recent efforts from our group to develop continuous processes based on the concept of countercurrent staging. This includes the use of countercurrent staged diafiltration for continuous protein formulation and the use of continuous countercurrent tangential chromatography for continuous steady-state product capture and purification.

8:50 Optimization of High-Throughput Antibody Purification Using Continuous Chromatography Matrices

Schmidt_PeterPeter Schmidt, PhD, Director, Recombinant Technologies Research, CSL Behring

Monoclonal antibodies are the fastest growing segment in the drug market. The development of mAbs requires purification of large numbers of variants with sufficient yield. However, established high-throughput purification strategies have been limited by the binding capacity of established affinity matrices. Our results show that matrices developed for continuous chromatography applications can help to overcome this limitation and increase the yield in high-throughput and lab-scale antibody purification.

Pall_Biotech 9:20 “A la carte” Menu for Process Intensification using Single-use Technologies and Continuous Bioprocessing

Collins_MikeMike Collins, Senior R&D Manager, Bioprocess R&D, Pall Biotech

Implementation of process intensification in the biopharmaceutical industry has led to increased process productivity, enabled process flexibility, improved process economics and reduced facility foot print. Adoption of single-use technologies is already partly responsible for this success. Switching from batch operations to continuous bioprocessing will leverage further the promises of process intensification.

9:50 Coffee Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

Overcoming Purification Challenges
Aqua EF

10:35

MsbA Structural Studies Using Novel Amphiphiles

Zhang_QinghaiQinghai Zhang, PhD, Associate Professor, Integrative Structural and Computational Biology, The Scripps Research Institute

MsbA is an inner membrane lipid A flippase and an essential ATP-binding cassette (ABC) transporter in gram-negative bacteria with homology to human multidrug resistance transporters. I will present our X-ray and EM structural studies of MsbA, which have been facilitated by the synthesis and characterization of novel stabilizing amphiphiles. The relevance of MsbA structures will be discussed in the context of a dynamic conformational pathway, thereby offering fresh insights into MsbA-mediated lipid A transport mechanism.

11:05 Detection and Assessment of Dilute Dosing Solutions of Potent Bispecific Molecules

Thomas_MelissaMelissa Thomas, PhD, Principal Scientist, Biologics – Protein Technologies, Amgen, Inc.

To ensure accurate dosing for Amgen’s BiTE therapeutic molecules, we need to assess drug product stability; however, detection and stability assessment of highly dilute protein solutions can be challenging. We have developed a sensitive HPLC-based method for detecting dilute solutions of proteins under simulated dosing conditions at concentrations of 100 ng/ml. This method has been used to evaluate and indicate potential liabilities of preclinical molecules, enabling selection and prioritization ahead of in vivo characterization.

11:35 Single-Step Purification of Intrinsic Protein Complexes for Functional Characterization in Saccharomyces cerevisiae Using Regenerable Calmodulin Resin: A Story of the ySet1C Enzyme-Substrate Network

Biggar_KyleKyle Biggar, PhD, Assistant Professor & Director, Carleton Functional Proteomics Facility, Biochemistry, Carleton University

The tandem affinity purification (i.e., TAP) method has been extensively used to purify native protein complexes under near physiological conditions in Saccharomyces cerevisiae. Our modification of this method provides an inexpensive single-step purification alternative to the traditional two step affinity purification of TAP-tagged proteins using only the calmodulin-binding peptide affinity tag. To demonstrate the effectiveness of our approach, we successfully purified and characterized the in vitro substrate preferences of the ySet1c methyltransferase complex.

12:05 pm Session Break

GenScript-CRO 12:15 Luncheon Presentation I: 
Accelerating Lead Identification Through Paramagnetic Bead Purification Technology

John Kawooya, PhD, Director, Biologics Optimization and Discovery Research, Amgen Inc.

HTP purification is central to screening lead panels of engineered biologics. Elimination of centrifugation and filtration steps through paramagnetic bead purification technology expedites this process faster than any other current technology on the market. Here, Amgen presents one of its latest parallel HTP paramagnetic bead innovation for antibody screening and purification. Some of Amgen’s magnetic platforms have been deployed and licensed to GenScript.  


GORE 12:45 Luncheon Presentation II: Improve Antibody Purification with Protein A Membrane Device

Barrett_BillBill Barrett, PhD, Product Specialist, Chromatography, Gore & Associates, Inc.

Protein capture has been identified as the number one bottleneck in the overall purification process of antibodies. The GORE™ Protein Capture Device with Protein A utilizes a unique membrane which provides high binding capacity at extremely short residence times. Productivity gains have been realized through the use of these devices in labs across the world. Results from current lab scale size devices and new test data will be presented on the next generation larger sizes.

 

1:15 Session Break


PLENARY KEYNOTE PANEL (Aqua Salon)

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PepTalk Perspectives: Point-Counterpoint Discussions

2:00 Plenary Keynote Introduction

Norman Packard, PhD, CEO, Daptics


2:10 Plenary Keynote Panel

Moderator:
Howard Levine, PhD, President and CEO, BioProcess Technology Consultants
Panelists:
George BadescuGeorge Badescu, PhD, Vice President, Scientific Affairs, Heidelberg Pharma AG
Cox ManonManon Cox, PhD, Co-Founder & CEO, NextWaveBio
Zhimei Du, PhD, Director, Bioprocess & Clinical Manufacturing, Merck
Paul Jorjorian, Vice President, BioProcess Sciences, Thermo Fisher Scientific
Marina Kirkitadze, PhD, Deputy Director, Head of Biophysics and Conformation Unit, Analytical R&D Biochemistry, Sanofi Pasteur, Canada
Stefan SchmidtStefan R. Schmidt, PhD, MBA, Head, Operations (COO), BioAtrium AG

 

3:05 Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

Innovating & Improving Purification Processes
Aqua EF

4:00 Chairperson’s Remarks

Qinghai Zhang, PhD, Associate Professor, Integrative Structural and Computational Biology, The Scripps Research Institute

4:05 Application of a Small EF Affinity Tag for Expression, Purification and SPR Studies of G Protein-Coupled Membrane Receptors

Yeliseev_AlexeiAlexei Yeliseev, PhD, Staff Scientist, Group Leader, LMBB, NIH, NIAAA

We developed a novel calcium-dependent EF-based affinity system that allows capture and high recovery of GPCR from dilute solutions containing detergents, salts and glycerol. The binding of the EF1 tag to the resin at these conditions is very strong that allows efficient purification without any loss of the target protein. The elution of the captured receptor is achieved by addition of EDTA, at very mild conditions that do not hinder the activity of this labile protein.

4:35 Structure and Protein-Protein Interactions in FAS and PKS 

Burkart_MikeMichael Burkart, PhD, Professor and Teddy Traylor Faculty Scholar, Chemistry and Biochemistry, University of California, San Diego (UCSD)

Fatty acid synthase (FAS) and polyketide synthase (PKS) pathways differ broadly in their identities and functional roles. Though the study of bacterial FAS has benefitted from decades of biochemical and structural investigations, PKSs have remained less understood, primarily due to challenges in protein expression and structural biology. Here we will discuss approaches to the structural and activity study of FAS and PKS utilizing the specificity conferred by protein-protein interactions. 

5:05 Establishing Innovative and Efficient Tool Boxes for Optimal and Scalable Processes for Recombinant Proteins

Shen_YuyiYuyi Shen, PhD, Associate Director, Process Development & Manufacturing, Bolt Biotherapeutics, Inc.

Innovative technologies evolve bioprocessing, and advance manufacturing in the pharmaceutical industry. The presenter will share case studies of successfully implementing innovative tools for process development and improvements for mAbs and complex recombinant proteins. The talk will discuss the major drive for innovative technologies and how to overcome key challenges of process integration and upgrades. The talk also provides insight into risk mitigation by balancing needs for quality, cost and speed.

5:35 A Universal Peptide-Tag System for Protein Purification and Analysis Based on Nanobody Technology

Rothbauer_UlrichUlrich Rothbauer, PhD, Professor, Natural and Medical Sciences Institute, University of Tübingen, and Co-Founder, ChromoTek GmbH

Single-domain antibodies - referred to as nanobodies - have emerged as an attractive alternative to traditional antibodies and became highly valuable tools for numerous bioanalytical and biotechnical applications. Here we present a novel nanobody-derived capture/detection system that enables fast and efficient isolation of epitope-tagged proteins from prokaryotic and eukaryotic expression systems. The high-affinity-binding and modifiable peptide tag of this system renders it a versatile and robust tool to combine biochemical analysis with microscopic studies.

6:05 - 7:00 Networking Reception in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

7:00 Close of Protein Purification and Recovery Conference