Antibodies Against Membrane Protein Targets Conference Part Two header


About This Conference:

As the pharmaceutical and biotech industries shift their attentions from small molecule therapeutics to biologics, an increased amount of attention is being paid to the prospect of membrane proteins as drug targets for antibodies and other protein scaffolds.  For these large target classes, biologics offer improved selectivity, an alternative for targets with known function that have not been amenable to small molecules and the potential for using antibodies for the targeted delivery of therapeutics.  However, for the field to advance, fundamental challenges of antigen quality, screening methodology, antibody engineering and target identification must be resolved.

This two-part meeting provides a forum in which discovery biologists and protein engineers can come together to discuss next generation strategies and technologies that will allow antibody-and alternate scaffold-based therapeutics directed against these target families to advance into the clinic and beyond.

The second conference, Generation, Preparation and Selection of Membrane Protein Targets, explores approaches to generating antigens of sufficient quality and purity to enable structural and modeling studies of target engagement and the associated screening and selection strategies used to isolate high-quality binders.



Thursday, October 9

11:30 am Registration


1:00 pm Plenary Keynote Program 
 

Chas BountraChas Bountra, Ph.D., Professor of Translational Medicine & Head, Structural Genomics Consortium, University of Oxford

Martin TolarMartin Tolar, M.D., Ph.D., Founder, President & CEO, Alzheon, Inc.

Andrew L. Hopkins, Andrew L. Hopkins, D.Phil, FRSC, FSB, Chair of Medicinal Informatics and SULSA Research Professor of Translational Biology, Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee


2:45 Refreshment Break in the Exhibit Hall with Poster Viewing


GENERATION OF FUNCTIONAL MEMBRANE PROTEINS

3:45 Chairperson’s Opening Remarks

Christopher Koth, Ph.D., Senior Scientist, Structural Biology, Genentech

3:55 Why VLPs for Generating Biologics to Cell Surface Proteins?

Mark Tornetta, Ph.D., Scientist, Molecular Discovery Technologies, Janssen Pharmaceuticals

Viral lipoparticles (VLP) present a great means to display cell surface targets. VLPs are either viruses or non-replicating viral particles. They contain the host cell’s membrane by way of budding, there in which the ability to capture the target protein in its natural conformation. This presentation will discuss VLPs as a format of ‘target display’ within the process of generating biological molecules to cell surface proteins.

4:25 High-Throughput Platforms for Expression of Bacterial and Eukaryotic Membrane Proteins

Renato Bruni, Ph.D., Head, Eukaryotic Membrane Protein Expression, New York Structural Biology Center

We describe here recent developments of two HTP platforms for the cloning and expression of integral membrane proteins. In the first one, prokaryotic membrane proteins are expressed in E.coli using a variety of expression vectors, growth conditions and purification methods. The second platform was developed for eukaryotic membrane proteins using mammalian cells for screening by fluorescence-detection size-exclusion chromatography and insect cells for expression and purification.

4:55 Creating Focused Mutant Libraries for Protein Engineering

Michael Drummond, Ph.D., Applications Scientist, Chemical Computing Group 

In protein engineering, efficient search and evaluation of excessively large sequence design space is challenging and requires multiple experimental rounds. Here we have developed a computational approach which predicts mutation probabilities for given residue sites in specified sequences. In assessing the probabilities at given residue sites, the sequence search space can be efficiently sampled to design and produce focused mutant libraries.

5:25 Coffee Break in the Foyer

5:40 Development of Engineered Crystallization Chaperones to Promote Membrane Protein Crystallization

Raquel Lieberman, Ph.D., Associate Professor, School of Chemistry & Biochemistry, Georgia Institute of Technology

We are developing for cost-effective non-covalent crystallization chaperones for membrane protein (MP) crystallization. The method is generalizable through insertion of a short epitope into a surface-exposed loop of a MP by site directed mutagenesis. Complexation and crystallization trials of representative chaperone-MP complexes are currently underway. In the long term, we hope these approaches will aid the community in solving structures of MPs of interest.

6:10 Optimization of Channels and Receptors for High-Throughput Screening and Antibody Development

Susmith Mukund, Senior Research Associate, Genentech

Membrane proteins are therapeutic targets for many diseases but are generally very difficult to express and purify. Considerable optimization is often required for downstream applications such as assay development, functional antibody screening and structural studies. This talk will provide a practical perspective on how to generate suitable purified membrane protein reagents in a drug development environment. Consensus ‘first-pass’ techniques for ion channel and receptor targets will be discussed, where functional antibodies or small molecule drugs are the desired outcome.

6:40 Close of Day

7:00 Dinner Short Courses*


*Separate registration is required


Friday, October 10

7:30 am Registration

8:00 Interactive Breakfast Breakout Discussion Groups 

This interactive session provides conference delegates and speakers an opportunity to choose a specific roundtable discussion group to join. Each group has a moderator to ensure focused discussions around key issues within the topic. This format allows participants to meet potential collaborators, share examples from their work, vet ideas with peers, and be part of a group problem-solving endeavor. The discussions provide an informal exchange of ideas and are not meant to be a corporate or specific product discussion.

Expression and Purification of Membrane Proteins

Renato Bruni, Ph.D., HeD, Eukaryotic Membrane Protein Expression, New York Structural Biology Center

  • Improving expression of eukaryotic membrane proteins using host and target mutagenesis approach 
  • Screening methods for selecting well-behaved MPs
  • Increasing the odds for successful purification and crystallization of MPs by way of chaperones

Preparation and Stabilization of Membrane Protein Targets for Antibody Production

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

  • Large scale expression of membrane receptor for antibody production
  • Detergents and lipids for stabilization of purified antigens.
  • Emerging technologies for stabilization and delivery of antigens

Can we Find Functional Antibodies to GPCRs and Ion Channels and Will They Make Good Drugs?

Mick Foley, Ph.D., Chief Scientific Officer, Biochemistry, AdAlta, Australia

  • Selection of antibodies; Immunization versus library selection
  • GPCR or ion channel format; overexpressed cells, membranes, or stabilized recombinant protein
  • How do we select for agonists, antagonist, inverse agonists etc?
  • Single domain, Fabs or whole IgG

PREPARATION OF MEMBRANE PROTEINS FOR ANTIBODY PRODUCTION

9:00 Chairperson’s Remarks

Rick Chu, Ph.D., Lead Research Investigator, Genzyme

9:10 Using Purified Membrane Proteins for Antibody Development: When and How

Christopher Koth, Ph.D., Senior Scientist, Structural Biology, Genentech

9:40 Stabilizing Membrane Protein and Membrane Protein Complex on Analytical Surface

Rick Chu, Ph.D., Lead Research Investigator, Genzyme

Traditionally, membrane protein binding assays rely on utilizing radioactive labeled ligands. In order to simplify membrane protein kinetics binding assay, purified membrane proteins, such as G-protein-coupled receptors (GPCRs), are captured on analytical surfaces and further stabilized by limited chemical crosslinking. This limited chemical crosslinking enables high quality label-free kinetics assays of membrane proteins via the same methods that are conventionally used for soluble proteins.

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

10:55 Challenges of Membrane Proteins Antigen Production for Antibody Generation

Ramkrishna (Ramu) Sadhukhan, Ph.D,. Senior Group Leader, Global Biologics, AbbVie


ASSAYS FOR DEVELOPING MABS AGAINST MEMBRANE PROTEIN TARGETS

11:25 Therapeutic Monoclonal Antibodies Targeting APJ Receptor

Krzysztof Palczewski, Ph.D., John H. Hord Professor and Chair, Department of Pharmacology, School of Medicine, Case Western Reserve University

The apelin receptor (APJ) is a G protein-coupled receptor (GPCR) widely expressed in various tissues, and it is associated with cardiovascular diseases and metabolic syndrome. A short peptide apelin is the only known APJ ligand, is rapidly degraded, and stable APJ agonists and antagonists with therapeutic potential are urgently needed. Monoclonal antibodies provide an attractive alternative strategy for targeting APJ therapeutically because they have significantly stability and high specificity. A strategy and molecular assays will be discussed toward developing therapeutically active mAb.

11:55 Characterization of a Nanobody Library Against Cannabinoid Receptor CB2

Alexei Yeliseev, Ph.D., Staff Scientist, Protein Biochemistry, LMBB, NIH

Cannabinoid receptor CB2 has become a prominent target for pharmaceutical drug development. We expressed and purified a full-length functional CB2, and optimized its stabilization in detergent micelles and lipid bilayers in a form of proteoliposomes and nanodiscs. A library of functional fold-specific camelid antibody fragments (nanobodies) was created, and their interaction with the receptor characterized by ELISA, G protein-activation assay, as well as by surface plasmon resonance and biolayer interferometry.

12:25 pm High-Throughput Microfluidic Platform for Functional Antibody Screening from Single Cells

Carl L. Hansen, Ph.D., President & CEO, AbCellera Biologics Inc.

AbCellera has developed a microfluidic-based platform for the high-throughput selection of monoclonal antibodies from single cells. This technology enables direct screening and characterization of antibody-secreting cells from any species using binding and functional cellular assays at a throughput of 400,000 cells per run.

12:55 Session Break 

1:05 Luncheon Presentation - Discovery of Rare Antibody Specificities to Difficult Targets Using High Content Screening of Avian Repertoires

Bill Harriman, Ph.D., CSO, Crystal Bioscience

Chickens are known to generate antibodies to epitopes on therapeutic targets that are highly conserved amongst mammals. These antibodies often demonstrate reactivity across multiple species, and are preferred when rodent or primate models of disease are anticipated. Using alternative immunization strategies we can enhance the prevalence of such clones, and by evaluating antibody profiles through a multi-parameter GEM screen of primary B cells, we can efficiently recover antibodies with desired biological activity and/or multispecies cross-reactivity.

1:45 Session Break 


ANTIBODY GENERATION, SELECTION AND SCREENING

1:55 Chairperson’s Remarks

Ralph Minter, Ph.D., Fellow, Technology, Antibody Discovery and Protein Engineering MedImmune, United Kingdom

2:00 Innovative Methods for the Generation of Nanobodies against Membrane Proteins

Jan Steyaert, Ph.D., Department Head, Structural Biology, Vrije University Brussels, Belgium

By rigidifying flexible regions and obscuring aggregative surfaces, nanobody complexes warrant conformationally uniform samples that are key to protein structure determination by X-ray crystallography. The elucidation of the first GPCR structure in its active state using a conformationally selective Nanobody demonstrates the power of the Nanobody platform to generate diffracting quality crystals of the most challenging targets including membrane proteins, and their complexes.

2:30 Phage Libraries and Their Application in Antibody Discovery Against Membrane Proteins

Johan Fransson, Ph.D., Principal Scientist, Janssen R&D

Antibody libraries displayed on phage offer a unique way to discover antibodies against virtually any protein. Specifically, cell panning strategies can be designed that enrich libraries with binders to a target cell line, while subtracting the vast majority of binders to non-target cells. This presentation will provide a perspective of the current state-of-the art selection schemes as well as insights into next generation phage libraries and their application.

3:00 Refreshment Break in the Exhibit Hall with Poster Viewing

3:30 Fluorescent Approaches for Screening and Characterizing Ligand and Antibody Binding to Membrane Proteins and Surface-Displayed Proteins in Yeast

Mark E. Dumont, Ph.D., Professor, Biochemistry & Biophysics, University of Rochester Medical Center

The genetic manipulability of yeast, coupled with genetic screening via flow cytometry, provides a powerful way of isolating rare variants with desired phenotypes from randomized libraries. We have used this approach to identify mutations of the endogenous yeast G protein coupled receptor Ste2p that specifically affect ligand binding, oligomeric state, and protein stability. We are also screening HIV envelope glycoprotein expressed at the yeast cell surface for variants exhibiting optimized antibody binding for vaccine development.

4:00 Activity-Based Screening of Antibodies to Cell Surface Targets

Ralph Minter, Ph.D., Fellow, Technology, Antibody Discovery and Protein Engineering MedImmune, United Kingdom

Target-led drug discovery is associated with a high attrition rate and also a high level of competition on each target. Our approach is to perform target-agnostic enrichment of antibodies on cells of interest and then to screen these antibodies for activity, prior to determining the target antigen. By following this phenotypic screening approach we have isolated many antibodies to novel disease-relevant targets, some of which will be described in more detail.

4:30 Single Domain Antibodies Against GPCRs and Ion Channels

Mick Foley, Ph.D., CSO, Biochemistry, AdAlta, Australia

The i-body is a human scaffold derived from information gained from the structure of the shark single domain antibody. The presence of a long CDR3 enables better access to GPCRs and ion channels than monoclonal antibodies. We have obtained high affinity single domain antibodies specific for the chemokine receptor CXCR4 that modulate β-arrestin signaling and block HIV infection. Binders to another GPCR LPA-1 and the ion channel TrpV4 have also been identified.

5:00 Close of Conference


Suggested Event Package

October 7

*Short Course: Targeting of GPCRs with Monoclonal Antibodies 

October 8-9: Antibodies Against Membrane Protein Targets – Part 1 Conference 

October 9-10: Antibodies Against Membrane Protein Targets – Part 2 Conference 


*Separate registration is required


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SPONSORSHIPS & EXHIBITS 

The exhibit hall was sold out in 2013, so please contact us early to reserve your place. To customize your sponsorship or exhibit package for 2014, contact:

Jon Stroup
Business Development Manager
781-972-5483
jstroup@healthtech.com 

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