Cambridge Healthtech Institute’s 7th Annual
Protein Purification and Recovery
January 21-22, 2015
The Protein Purification and Recovery conference explores how experts are innovating processes to achieve purity, and provides an overview of the current tactics and technologies for optimizing protein purification in the effort to reach consistency and quality. Case studies will be presented that illustrate the tenacity needed to maximize purity and yield, while minimizing purification steps. The agenda addresses process development and issues of scale, along with purifying challenging proteins, such as membrane proteins. The meeting also addresses purification of proteins in different expression systems, including mammalian, bacterial and insect cells.
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TUESDAY, JANUARY 20
1:30 pm Conference Registration
2:00 BuzZ Session
3:00 Refreshment Break in the Exhibit Hall with Poster Awards
3:45 BuzZ Session B
4:30-5:00 Short Course Registration
5:00-8:00 Dinner Short Courses More Details >>
WEDNESDAY, JANUARY 21
7:30 am Conference Registration and Morning Coffee
8:15 Chairperson’s Opening Remarks
William Gillette, Ph.D., Senior Scientist, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research
8:20 The Distinct Issues of Membrane Proteins versus Soluble Proteins
Robert Stroud, Ph.D., Professor, Biochemistry/Biophysics and Pharmaceutical Chemistry; Director, Membrane Protein Expression Center, University of California at San Francisco (UCSF)
Membrane proteins present distinct issues versus soluble proteins; pure, homogeneous and stable in solution. Eukaryotic proteins need expression in eukaryotic cells. These include yeast, and insect cells. Human proteins, to be expressed for antibody preparation, require proper tailoring and glycosylation; and can often be achieved only in human cells. Antibody partners can be the basis for proof of principle therapeutics, and for helping to purify, assay, and validate membrane proteins’ mechanisms.
9:00 Expression of Transmembrane Proteins in Yeast for Genetic, Biochemical and Structural Studies
Mark E. Dumont, Ph.D., Professor, Biochemistry and Biophysics, University of Rochester Medical Center
The difficulty of expression, solubilization, and purification of transmembrane proteins, particularly those from eukaryotes, constitutes a significant barrier to understanding their mechanisms. The use of a system based on expression of membrane proteins from various sources in baker’s yeast has allowed the use of genetic approaches for functional characterization and stabilization of expressed proteins, while facilitating purification of transmembrane enzymes, transporters, and receptors for biochemical studies and x-ray crystallographic structure determination.
9:30 A Novel Strategy to Express and Purify Highly Stable and Functional GPCR Signaling Complexes
Arun Shukla, Ph.D., Assistant Professor, Medicine, Duke University
Structural characterization of GPCR signaling complexes remains a very challenging but extremely important task. Molecular visualization not only reveals fundamental mechanism of assembly and functioning of such complexes but also provides a unique framework for drug discovery. We present a novel strategy to assemble a highly stable and functionally competent GPCR signaling complex in-cellulo and its subsequent purification for direct structural characterization.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing
10:50 GPCR Purification: Protein Engineering and Process Optimisation are Key Elements for Successful GPCR Crystallisation
Markus Koglin, Ph.D., Associate Director, Protein Engineering, Heptares Therapeutics
The generation of Stabilised G-protein Coupled Receptors (StaRs) has opened new routes in the purification procedures for GPCRs. Here we demonstrate that protein quality is drastically improved with increasing thermostability. StaR generation alone normally does not provide suitable material for crystallisation. The combination of StaR technology, systematic construct design, and careful optimisation of the purification process delivers a powerful approach to generate protein with suitable crystallisation qualities.
11:20 Strategies for Detergent Stabilization and Large-Scale Affinity Purification of the Functional G Protein-Coupled Receptors
Alexei Yeliseev, Ph.D., Staff Scientist, Group Leader, LMBB, NIAAA, National Institutes of Health (NIH)
Human cannabinoid receptor CB2, a G protein-coupled receptor involved in regulation of immune response, is an important target for pharmaceutical drug development. For high resolution structural studies, milligram quantities of pure, homogenous and functional protein are required. We expressed the functional CB2 receptor in E. coli, and optimized its purification by tandem affinity chromatography. An efficient strategy for stabilization of the receptor in detergent micelles and reconstituted into lipid bilayers was developed.
11:50 Overcoming Difficult to Express Proteins: Cell-Free Additives for Solubilizing and Characterizing Membrane Proteins
Matthew Coleman, Ph.D., Deputy Group Leader, Molecular Toxicology, Lawrence Livermore National Laboratory; Senior Scientist, University of California at Davis
We have developed cell-free methods for producing membrane proteins, which are inherently difficult to obtain. These include nanolipoprotein particles (NLPs), telodendrimers, which can be combined with NLPs or lipids, and amphipathic peptides. I will further discuss how we use the three different additives and their ability to support membrane protein solubility as well as function. These processes are easily adapted to high-throughput technologies for feeding the membrane structural biology pipeline.
12:20 pm Sponsored Presentation (Opportunity Available)
12:50 Session Break
1:00 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
2:00 Chairperson’s Remarks
Matthew Coleman, Ph.D., Deputy Group Leader, Molecular Toxicology, Lawrence Livermore National Laboratory, and Senior Scientist, University of California at Davis
2:05 Resolving Protein Purification Challenges for Therapeutic Antibody Development
Julie Q. Hang, Ph.D., Senior Scientist, Protein Chemistry, Genentech, Inc.
Purification technology is fundamental in antibody drug discovery. Maintaining biological and biophysical features of antigens is critical to generate antibodies with high diversity and specificity. Various antigen forms and orthologs are required for antibody screening and cross-species affinity. With increasing numbers of new formats of antibody fragments as therapeutic proteins, there are more technical demands in purification and characterization. This talk will present case examples in dealing with target-specific challenges.
2:35 Development of an Industrially Relevant DSP Platform Process for a Bispecific Antibody, the κλ-Body
Jean-François Depoisier, Head, Downstream Processing Unit, NovImmune SA
In order to exploit novel mechanisms of action and achieve superior clinical efficacy, NovImmune has developed a novel bispecific antibody format, the κλ-body that has a molecular structure identical to a fully human monoclonal antibodies. The talk will address the challenges in developing an industrially relevant purification process for this innovative bispecific antibody format. The strategy for identifying optimal work space based on a broad range scanning of process parameters and novel purification technologies will be presented.
3:05 BEAT Bispecific Antibody: Development of a Process with a Platform Approach
Sonia Letestu, Process Development Engineer, DSP, Glenmark Pharmaceuticals S.A.
The BEAT® is a novel bispecific antibody techonology developed by Glenmark Pharmaceuticals. The scFv-FAB BEAT® format is a combination of a Fab arm and an ScFv arm with a fully functional Fc part, conserving key IgG properties such as thermostability, the potential for effector function and antibody-like pharmacokinetics. The presentation will present a new Bispecific technology specically engineered in order to allow an efficient impurity removal in one single step.
3:35 A New Alkali Stable Adsorbent for Antibody Purification
Steve Burton, Ph.D., CEO, ProMetic BioSciences Ltd.
4:05 Refreshment Break
4:25 Plenary Keynote Session
5:45-7:00 Reception in the Exhibit Hall with Poster Viewing
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