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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

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

Purifying Antibodies

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

Basar 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.

3:15 Sponsored Presentation (Opportunity Available)

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

Purifying Antibodies (Cont.)

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 MsbA Structural Studies Using Novel Amphiphiles

Qinghai 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.

5:30 Close of Day

5:30 - 5:45 Short Course Registration


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

Click here for more details.

*Separate registration required

WEDNESDAY, JANUARY 16

7:45 am Registration and Morning Coffee

Continuous Technology for Antibody Purification

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

Andrew 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

Peter 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.

9:20 Sponsored Presentation (Opportunity Available)

9:50 Coffee Break in the Exhibit Hall with Poster Viewing

Overcoming Purification Challenges

10:35 Development and Optimization of Challenging Unit Operations with Line of Sight to Manufacturing for a Shear Sensitive, Aggregation Prone, & Low pI Monoclonal Antibody

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

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

Melissa 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

Kyle 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

12:15 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:15 Session Break


2:00 PLENARY KEYNOTE PANEL

Click here for more details.

PepTalk Perspectives: Point-Counterpoint Discussions

Moderator:
Howard Levine, PhD, President and CEO, BioProcess Technology Consultants






Panelists:
Zhimei Du, PhD, Director, Bioprocess & Clinical Manufacturing, Merck






Lorenz Mayr, PhD, CTO, GE Healthcare Life Sciences




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

Innovating & Improving Purification Processes

4:00 Chairperson’s Remarks

Basar 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

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

Alexei 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 A Virtual QC Platform for High-Throughput Protein Purification

Philip E. Hass, Senior Scientific Manager, Genentech, Inc.

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

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

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

Ulrich 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

7:00 Close of Protein Purification and Recovery Conference

* The program is subject to change without notice, due to unforeseen reason.

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