January 18-22, 2016 | Town and Country Resort Hotel | SAN DIEGO, CA 
January 18-22, 2016 | Town and Country Resort Hotel | SAN DIEGO, CA 

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Cambridge Healthtech Institute’s 2nd Annual
Membrane Proteins
A Valuable Resource and Target
January 19-20, 2016


Membrane proteins are the gateways to the cell and are valuable drug targets. For researchers to design better-targeted drugs, they need to know their structure and functional characteristics. Cambridge Healthtech Institute’s 2nd Annual Membrane Proteins conference addresses the strategies and solutions for their extraction, expression and purification, and features case studies showcasing their value as a drug target. Join the in-depth exploration of how to obtain functional membrane proteins, and learn more about this important protein class.

Final Agenda

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TUESDAY, JANUARY 19

1:00 pm Conference Registration


PURIFYING MEMBRANE PROTEINS

2:00 Chairperson’s Opening Remarks

William Gillette, Ph.D., Senior Scientist, Protein Expression Laboratory, Cancer Research Technology Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research (FNL)


KEYNOTE PRESENTATION

2:05 Making Water-Soluble Integral Membrane Proteins in vivo Using an Amphipathic Protein Fusion Strategy

Matthew P. DeLisa, Ph.D., William L. Lewis Professor, Chemical & Biomolecular Engineering, Cornell University

Here we devise a general strategy for in vivo solubilization of IMPs in structurally relevant conformations without the need for detergents or mutations to the IMP itself. This technique, called SIMPLEx (solubilization of IMPs with high levels of expression), allows the direct expression of soluble products in living cells by simply fusing an IMP target with truncated apolipoprotein A-I, which serves as an amphipathic proteic ‘shield’ that sequesters the IMP from water and promotes its solubilization.


⊲ Featured Presentation
2:45 Expression and Sample Preparation of Membrane Proteins for Structure Determination by NMR

Stanley Opella, Ph.D., Professor, Chemistry and Biochemistry, University of California, San Diego

The advantages of heterologous expression of proteins in bacteria include the ability to make relatively large amounts and the ready incorporation of stable isotopes. The use of a hydrophobic fusion protein enables the sequestration in inclusion bodies to avoid damaging the cell membrane.

3:15 Refreshment Break in the Exhibit Hall with Poster Awards

4:00 Selexis SUREcode™: De-Risking Research Cell Bank Generation with Comprehensive Genomic Analysis

Pierre-Alain Girod, Ph.D., CSO, Selexis

RCBs are cell populations with neither identical genomes nor single integration sites even when selected from single isolated clones. The inherent genomic instability of CHO-K1 leads to the appearance of mixed populations that can result in decreased manufacturability. Selexis’s proprietary SUREcode™ bioinformatics, based on Next Generation Sequencing of entire genomes, provides unique and detailed genomic maps of cell populations. It is used to fully characterize integration sites and transgene sequences as well as provide detailed genomic information that reduces and mitigates risk while manufacturing recombinant protein drugs.

4:30 It Takes Two to Tango—Structure/Function Studies Yield the Dance of the Permease

H. Ronald Kaback, M.D., Distinguished Professor, Physiology, University of California, Los Angeles

Lactose permease (LacY) catalyzes translocation of a galactoside and an H+ across the membrane. X-ray structures, and structure/function studies reveal that: (1) LacY utilizes an alternating access mechanism; (2) sugar binding involves induced fit; (3) Active transport does not involve a change in KD for sugar on either side of the membrane, but the pKa decreases markedly. (4) Transport is driven chemiosmotically, and ∆µ̃H+ acts kinetically to accelerate the process.

5:00 Strategies for High Yield Affinity Purification of Functional G Protein Coupled Receptor from Detergent Solutions

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

Human cannabinoid receptor CB2, a G protein-coupled receptor involved in regulation of immune response, is an important target for pharmaceutical drug development. We expressed the functional CB2 receptor in E. coli, and optimized its purification by tandem affinity chromatography using novel affinity resins StrepTactin XT Superflow and EF2 Ca-calbindin-based resin. Examples of successful purification and efficient recovery (over 80%) of CB2 from dilute detergent-containing solutions will be presented.


5:30-5:45 Short Course Registration

5:45-8:45 Dinner Short Courses


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WEDNESDAY, JANUARY 20

8:00 am Conference Registration and Morning Coffee


PURIFYING ANTIBODIES

8:30 Chairperson’s Remarks

Hong Li, Ph.D., Principal Scientist, Purification Process Development, Merck

8:35 Purification of Common-Light-Chain Bispecific Antibodies

Juergen Nett, Ph.D., Associate Director, 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. A panel of bispecific constructs are then generated that bind to each target with high affinity and exhibit favorable biophysical properties similar to traditional therapeutic antibodies.

9:05 A Simple, Robust Two-Column Purification Platform for Antibody Fragment Manufacturing

Green Guihang Zhang, Ph.D., Director, Protein Sciences, ImaginAb, Inc.

Minibody and cys-diabody are unique types of antibody fragments developed for clinical imaging. Purification of minibodies and cys-diabodies has proved to be challenging due to their high aggregation and low-pH sensitivity. We have recently developed a simple and robust two-column purification platform for manufacturing minibody and cys-diabody for clinical applications with high purity and stability. The same platform may be used for other types of antibody fragment purification.

9:35 Presentation to be Announced


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

10:50 Developing a Purification Platform for FAB

Jiansheng Wu, Ph.D., Senior Scientist, Protein Chemistry, Genentech, Inc.

FAB has gained more attention in recent years due to its broad applications in therapeutics and diagnostics. Generating grams level of FAB from E.coli cell pellets is important for many preclinical studies. We evaluated several resins for FAB purification and developed a robust purification platform to obtain up to 20 grams of FAB from E. coli cells.

11:20 Downstream Purification Case Studies

Yun Bai, Ph.D., Director, Process Development, Ambrx, Inc.

11:50 Aggregation Challenges during the Purification Development of a Low pl Monoclonal Antibody

Hong Li, Ph.D., Principal Scientist, Purification Process Development, Merck

12:20 pm Presentation to be Announced


12:50 Session Break

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


PURIFICATION STRATEGIES

2:00 Chairperson’s Remarks

Jiansheng Wu, Ph.D., Senior Scientist, Protein Chemistry, Genentech, Inc.

2:05 Purification of Secretory IgA from Lemna minor (Duckweed)

Daniëlle van Wijk, Ph.D., Lead Scientist / Project Leader, Down Stream Processing, Synthon Biopharmaceuticals B.V.

Here we present the duckweed Lemna minor as a promising platform for production of SIgA antibodies. The production process comprises (1) expression of SIgA in stably-transformed duckweed; (2) extraction of SIgA by disruption of the plant material; (3) removal of naturally abundant impurities by acidic precipitation; (4) clarification by depth filtration and TFF; (5) purification by affinity chromatography followed by polishing steps; and (6) formulation in a stable buffer.

2:35 Protein Purification for the RAS Initiative at the Frederick National Lab

William Gillette, Ph.D., Senior Scientist, Protein Expression Laboratory, Cancer Research Technology Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research (FNL)

One of the major goals of the RAS Initiative at the Frederick National Lab is to deepen the knowledge of the RAS proteins. In support of that effort, the Protein Expression Laboratory has focused its effort on the purification of KRAS4b, its oncogenic mutants, and interacting protein partners in a multidisciplinary effort. Biophysical characterization and functional assays of the proteins will be discussed.

3:05 The Use of Affinity Tags to Overcome Obstacles in Recombinant Protein Expression and Purification

Jian-Ping Jin, M.D., Ph.D., Professor and William D. Traitel Endowed Chair, Physiology, Wayne State University School of Medicine

To overcome obstacles in the expression and purification of recombinant proteins are of importance in research and industrial applications. A strategy is the use of affinity tags or carrier peptide. Strategies have also been developed to remove the tag after purification and obtain native protein and peptide products. There are unsolved problems and imperfect applications. The pros and cons of current approaches will be discussed for improvement and future development.

3:35 Refreshment Break in the Exhibit Hall with Poster Viewing


STREAMLINING PURIFICATION PROCESSES

4:30 Crystallization as a Tool for Scalable, Efficient Purification Techniques in Early Process Step for Purifying Recombinant Proteins

Partha Hazra, Ph.D., Chief Scientific Manager, Research, Biocon

Crystallization/precipitation can be a simplified, efficient and cost effective tool for separation of both process and product related impurities in processing of recombinant therapeutic proteins. I will present a Case Study comparing traditional Capture chromatography and Crystallization/precipitation steps, and will cover the advantages of one over the other when manufacturing at large scale. Comparative quality observed in the scale up experiences also will be covered in the case study.

5:00 pH-Dependent Sharkbodies for Affinity Purification of Biologics

Harald Kolmar, Ph.D., Professor and Head, Applied Biochemistry, Chemistry, Technical University of Darmstadt

We have established a platform for generation of shark-based antibody domains that display pH-dependent target protein binding. To this end, a master library of sharkbodies with random histidine-enriched binding loops was generated that are displayed on yeast cells and screened by high-throughput FACS. Due to their high inherent stability and pH-dependent binding characteristics they are excellently suited for affinity chromatography purification of biologics, particularly for non-antibody protein therapeutics.

5:45 BuzZ Session B

Join your peers and colleagues for interactive roundtable discussions.

6:30-7:30 Reception in the Exhibit Hall with Poster Viewing

7:30 Close of Conference


Day 1 | Day 2 | Download Brochure