As the pharmaceutical and biotech industries increasingly shift attention to biologics, much more attention is being paid to the prospect of membrane-bound proteins as drug targets for antibodies and other protein scaffolds. For the large GPCR and ion channel target classes, biologics offer improved selectivity, an alternative for targets with known function that have not been amenable to small molecule drugs and the potential for using antibodies for the targeted delivery of therapeutics. However, for the field to advance, fundamental challenges in optimizing antigen quality and presentation, discovery methodologies, protein engineering and target identification must be resolved.

The two-part Antibodies Against Membrane Protein Targets 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 in the set offers an examination of state-of-the-art approaches for the expression of high quality membrane protein antigens and antibody generation, then explores selection and screening strategies that can be applied to discover binders with functional activity against GPCR and ion channel targets.

Final Agenda

Thursday, September 27

11:50 am Conference Registration Open

12:20 pm Plenary Keynote Program

2:00 Refreshment Break in the Exhibit Hall with Poster Viewing

Discovery Of Functional Antibodies

2:45 Welcome Remarks

Kent Simmons, Senior Conference Director, Cambridge Healthtech Institute

2:50 Chairperson’s Opening Remarks

Iain D. G. Campuzano, Principal Scientist, Discovery Attribute Sciences, Amgen

2:55 Droplet-Microfluidics for the Discovery of Antibodies Binding or Modulating Receptors on Target Cells

Christoph Merten, PhD, Group Leader, Microfluidics, European Molecular Biology Laboratory (EMBL), Germany

Droplet microfluidics has become a powerful tool for antibody screening. After giving a general overview on the technology, I will introduce our new 2-cell platform, facilitating assays involving two different cell types. This allows screening hybridoma cells or even primary plasma cells (from mice and humans) for the secretion of antibodies binding or modulating membrane receptors on specific target cells. Approximately 100,000 antibodies can be screened in a single experiment in less than 24 hours.

3:25 Strategies for Antibody Discovery to Integral Membrane Proteins

Robert Pejchal, PhD, Associate Director, Antibody Discovery, Adimab LLC

Discovery of antibodies that target integral membrane proteins at the cell surface represent a unique challenge. Using yeast presentation of large synthetic human antibody libraries, Adimab has developed methods for isolation of target specific binders utilizing whole cells selection, NGS, and high-throughput screening. Considerations for utilization of solubilized protein are also discussed.

3:55 Presentation to be Announced

4:10 Sponsored Presentation (Opportunity Available)

4:25 Refreshment Break in the Exhibit Hall with Poster Viewing

5:00 Conformational Display Selection of Functional Antibodies against GPCRs

John Burg, PhD, Head, Biochemistry, Ab Initio Biotherapeutics

G protein-coupled receptors (GPCRs) are critical signaling mediators for both endogenous ligands and therapeutics. By combining next-generation synthetic antibody libraries with novel selection techniques, we have enabled the facile discovery of functional antibodies against GPCRs. Our selections have yielded antibodies that recognize native extracellular GPCR epitopes, with the ability to modulate receptor signaling. Our technology opens the door to routine discovery in this important new class of biologics.

5:30 Generation of Positive Allosteric Modulators of TRPV1 Using Affimer Reagents

Darren Tomlinson, PhD, Associate Professor, University of Leeds, United Kingdom

Affimers are scaffolding proteins that constrain variable regions for molecular recognition and can be used as alternatives to antibodies in many applications. In this talk, I will focus on the isolation of Affimer reagents against TRPV1, an ion channel implicated in pain disorders. The reagents act as positive allosteric modulators, and we have mimicked the interaction with small molecules through in silico drug design based on a co-crystal structure.

6:00 Employing High Throughput Single B Cell Cloning Platforms to Derive Functional Therapeutic Antibody Candidates against Membrane Protein Targets

Jason Damiano, PhD, Director, Antibody Therapeutics, Achaogen

Integral membrane proteins remain a substantial untapped pool of targets which can mediate innovative approaches to treat human disease. Utilizing antibody-based therapeutics as a modality to effectively modulate the function of this target class offers many advantages, but their discovery is often challenging. Through the utilization of technologies which enable upfront functional interrogation of antibodies secreted by individual B cells, we have rapidly identified therapeutic candidates against disease-relevant transmembrane proteins.

6:30 Dinner Short Course Registration

9:30 Close of Day

Friday, September 28

7:00 am Registration Open

7:30 Interactive Breakfast Breakout Discussion Groups

Grab a cup of coffee and join a breakout discussion group. These are informal, moderated discussions with brainstorming and interactive problem solving, allowing participants from diverse backgrounds to exchange ideas and experiences and develop future collaborations around a focused topic. Details on the topics and moderators are available on the conference website.

Antibody Generation

8:30 Chairperson’s Remarks

Christopher Murawsky, PhD, Principal Scientist, Amgen

8:35 Raising Antibodies to GPCRs: A Retrospective Analysis of Successes and Challenges at BMS

Dan Rohrer, PhD, Senior Director, Biologics Discovery, Bristol-Myers Squibb

Raising antibodies to GPCRs, especially those that interact with native GPCR conformations, is a technical challenge that has been addressed by every facet of the BMS antibody discovery process. Presentation of stable and native structures is key, but many other variables also play a role in successful campaigns, such as adjuvant use and immunization schemes, host animals of differing immune response, and novel screening methods. We will review several examples and discuss some of the lessons learned by our team.

9:05 Antibody Generation for Membrane Proteins and Complex Epitopes

Christopher Murawsky, PhD, Principal Scientist, Amgen

Many of the most therapeutically interesting targets are multi-pass or multi-subunit membrane proteins that have nominal surface area exposed that can serve as epitopes for antibody binding. The extracellular regions may interact to form complex, non-linear epitopes that are required for activity and are the targets of function-modifying antibodies. We present case studies describing strategies to discover antibodies to these targets, focusing on antigen engineering, generation of large and diverse antibody repertoires and high throughput screening.

9:35 Improving Antibody Discovery against Challenging Targets

Rajesh Vij, Senior Scientific Researcher, Antibody Engineering, Genentech

The generation of large and diverse panels of monoclonal antibodies against multi-transmembrane or integral membrane proteins has proven challenging, thus hindering the discovery of rare functional clones. Focusing on a case study, we will discuss how antigen format, immunization scheme, and downstream expression and screening strategies can be optimized to identify antibodies with novel activities.

10:05 Coffee Break in the Exhibit Hall with Poster Viewing and Poster Competition Winner Announced

10:45 Cell Membrane Preparation to Enable Immunization, Antigen-Specific B-Cell Sorting and Screening for Antibodies against Cell Surface Targets

Habtom Hapte, PhD, Principal Scientist, Biotherapeutics Discovery, Boehringer Ingelheim

Here we report a strategy that resulted in identification of antigen-specific B cells using membrane prep as bait and a membrane prep based Meso-Scale Discovery (MSD) screening. Single B-cell sorting using membrane prep as bait generated 118 target-1-specific hits. This strategy will enable us to generate immune responses against targets that are difficult to express and purify using membrane prep based immunization, sort and screen.

11:15 So Long Llamas: Rapid Discovery of Synthetic GPCR-Targeted Nanobodies

Conor McMahon, PhD, Postdoctoral Fellow, Biological Chemistry and Molecular Pharmacology, Harvard Medical School

Single-domain camelid antibody fragments (‘nanobodies’) are broadly used as tools for biological research and have therapeutic potential. However, nanobody discovery typically requires animal immunization, presenting a host of barriers to their production. We developed a fully synthetic yeast displayed nanobody library, which circumvents animal immunization and also allows rapid isolation of conformation-specific binders. From this library, we generated a range of functional nanobodies targeting GPCRs, including conformational stabilizers and ligands.

11:45 Sponsored Presentation (Opportunity Available)

12:15 pm Session Break

12:25 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:15 Refreshment Break in the Exhibit Hall with Poster Viewing

Generation And Optimization Of Membrane Protein Antigens

1:55 Chairperson’s Remarks

Dan Rohrer, PhD, Senior Director, Biologics Discovery, Bristol-Myers Squibb

2:00 Advances in Target Expression and Display for Discovery of Biological Therapeutics

Ivan Correia, MBA, PhD, Research Fellow, Head, Global Protein Sciences, AbbVie Bioresearch Center

Generation of specific immune response to a target protein is dependent on correct fold and conformation state of the antigen and must represent its natural state in vivo. Other important considerations include post-translational modifications and genetic polymorphism. Progress is continuously being made, and in this presentation, we introduce new perspectives and advances. We highlight many options available and successfully applied to a large number of target proteins.

2:30 Beyond Detergents: Nanoencapsulation of Membrane Proteins for Drug Discovery

Tim Dafforn, PhD, Professor, Biotechnology, University of Birmingham, United Kingdom

For more than 40 years, the only way to extract membrane proteins from cells was to use a detergent. However, this approach was far from successful, producing samples with low stability and activity. The recent emergence of nanoencapsulation systems like MSP and SMALP now provide membrane protein samples with high stability. In this talk, I will summarize these approaches and new developments that have occurred in the SMALP field.

3:00 Expression and Screening of Human Integral Membrane Protein Targets

Nicola Burgess-Brown, PhD, Principal Investigator, Structural Genomics Consortium, University of Oxford, United Kingdom

The SGC promotes research advancement through our open access policy, and in the absence of IP. Globally, we have solved more than 2000 human protein structures and 10 novel integral membrane proteins (IMPs). Although we have made a significant contribution to structural biology and protein production for functional studies, IMPs and protein-protein complexes still remain a challenge to produce. Here, I present our established approaches for eukaryotic expression and screening IMPs using baculovirus/insect cells and BacMam technology.

3:30 Emerging Strategies for Membrane Protein Presentation in Antibody Discovery

Pawel Dominik, PhD, Senior Scientist, Cancer Immunology Discovery, Pfizer

Phage display enables the discovery of high affinity antibodies to a variety of antigens. While discovering antibodies has become routine for most proteins, it is still challenging for transmembrane proteins. Nanodiscs have emerged as powerful tools to study membrane proteins in a more natural lipid environment. By combining phage display and nanodiscs, we improved our ability to discover antibodies to multi-pass transmembrane proteins. This approach expands the toolbox for the rapid discovery of therapeutic antibodies.

4:00 Close of Conference