The field of protein engineering is at an exciting point in its development, with new generations of therapeutic antibodies now reaching the market, great advances in protein science and a body of clinical evidence that can be used to inform the development
of safe, highly effective therapies for unmet medical needs. The PEGS Engineering Antibodies conference explores the state of the art in technologies used by protein engineers working at the discovery and design stages to quickly
and efficiently craft biotherapeutics directed at the most elusive targets and biological functions.
Final Agenda
Recommended Short Courses*
SC13: Phenotypic Screening Applications and Technologies - Detailed Agenda
SC14: Overcoming the Challenges of Immunogenicity Assays, Risk Assessment and Meeting Regulatory Requirements - Detailed Agenda
*Separate registration required
WEDNESDAY, MAY 3
7:30 am Registration and Morning Coffee
8:30 Chairperson’s Remarks
Robin Barbour, Head, Antibody and Assay Development, Research, Prothena Biosciences
8:40 KEYNOTE PRESENTATION: New High-Throughput Model for Optimizing Antibody Affinities
Frederick Alt, Ph.D., Charles A. Janeway Professor of Pediatrics, Boston Children’s Hospital; Professor of Genetics, Harvard
Medical School; Director, Program in Cellular and Molecular Medicine, Boston Children’s Hospital
We will describe our new approach to utilize natural primary and secondary mechanisms of antibody optimization in vivo to improve therapeutic antibodies. We use genetically
modified ES cells to rapidly generate chimeric mice that extensively diversify a given therapeutic antibody. Following immunization, variant therapeutic antibodies with superior properties to the prototype can be selected. We validated this system
by substantially improving the antigen binding affinity of an existing therapeutic antibody.
9:10 Novel Mechanism of Antigen Recognition by Extremely Specific Synthetic Antibodies
Shohei Koide, Ph.D., Professor, Biologics Design, Langone Medical Center, New York University
Creating molecular recognition interfaces that discriminate subtle chemical differences remains a major challenge in protein engineering. We have generated synthetic antibodies against histone tails that cleanly discriminate the difference of a single
methyl group, arguably the smallest difference in antigens. Crystal structures and mechanistic studies revealed a surprising mechanism underlying their exquisite specificity, which has led to the design of new antibody formats suitable for achieving
exceptional specificity.
9:40 Electron Microscopy Structures of Dual Variable Domain Immunoglobulin (DVD-Ig1) and its Complex with Target Antigen
Paul Matsudaira, Ph.D., Professor, Dept. of Biological Sciences, National University of Singapore
A dual variable domain immunoglobulin, DVD-Ig1, forms 1:2 and 2:2 complexes with its antigen, TNFα. Our negative stain TEM studies show that DVD-Ig1 displays flexibility with different angles between the Fab arms and dynamics by the two Fab’s
outer domains: without TNFα (DVD-Ig1 alone) and with TNFα (DVD-Ig1/TNFα complexes). In addition, the structures of these molecules and complexes were studied using single particle cryoEM and preliminary findings will be discussed.
10:10 Coffee Break in the Exhibit Hall with Poster Viewing
10:55 Droplet Microfluidics in High-Throughput Antibody Discovery and Vaccine Development
Christoph Merten, Ph.D., Group Leader Microfluidics, European Molecular Biology Laboratory (EMBL)
We have developed droplet-based microfluidic platforms for the discovery of therapeutic antibodies. The technology allows the direct screening of >1 million primary, non-immortalized plasma cells (optionally from humans) in a single experiment
and also facilitates assays for the effect of antibodies on target cells (e.g. modulating GPCRs). In a complementary approach we use the technology to derive vaccine candidates starting with neutralizing antibodies against pathogens such as HIV.
11:25 Germline-Encoded Neutralization of a Staphylococcus Aureus Virulence Factor by the Human Antibody Repertoire
Andy Yeung, Ph.D., Associate Research Fellow, Rinat-Pfizer
We employed single-B cell cloning, phage display, high-throughput sequencing, epitope mapping, and crystallography to characterize in detail the humoral immune response to the staphylococcal protein IsdB. We show that in all donors a heavily
biased use of two immunoglobulin heavy chain germlines generated ultra-high affinity neutralizing antibodies. Interestingly, the binding is primarily driven by the germline-encoded CDR-H2, with a binding mechanism nearly identical for
each antibody derived from different donors.
11:55 Antibody Specificity Profiling by Combining NGS with a Plug-and-(Dis)play Hybridoma Platform
Sai Reddy, Ph.D., Assistant Professor, Biosystems Science and Engineering, ETH Zurich
In this presentation, I will show how we are combining this NGS-based analysis with a novel mammalian hybridoma display platform for antibody screening and discovery. Specifically, we use NGS to identify candidate antigen specific clones from
immunized repertoires, we then integrate these antibody clonal libraries into our hybridoma platform using CRISPR-Cas9 genome editing. Flow cytometry is then used to screen and isolate antigen-specific antibodies.
12:25 pm CHO Cell Line Engineering for Enhanced Productivity and Stability
Pierre-Alain Girod, Ph.D., CSO, Selexis
Stable, high quality production cell lines secreting optimal levels of recombinant protein require stable integration of the recombinant DNA, elevated gene transcription, optimized secretion and metabolic machinery to handle the increased
protein loads along with cellular phenotypic stability. Using the extensive transcriptomic and FISH-RNA/DNA data we have generated for our CHO-K1 cell line (CHO-M), we will describe how we are significantly boosting production capabilities
and cell line stability of our CHO-M cell line.
12:55 Luncheon Presentation I: The Trianni Mouse: Best-In-Class Technology for Human Antibody Discovery
David Meininger, Chief Business Officer, Trianni, Inc.
The Trianni Mouse is the only human transgenic antibody discovery platform offering a complete heavy, kappa and lambda repertoire in a single organism. Sequences of the variable domain exons are human while all genetic machinery are of mouse
origin. The platform is seen as best-in-class by multiple Big Pharma and other licensees subsequent to extensive validation and benchmarking. Additional strains in development include Plasma Ig, Autoimmune/All Epitope and a “true”
Bispecific.
1:25 Luncheon Presentation II: HuMab Chickens: The Next Generation Antibody Discovery Platform
Bill Harriman, Ph.D., CSO, Crystal Bioscience
Transgenic rodents producing human sequence antibodies are widely accepted as a reliable source of therapeutic candidates. However, their repertoires are limited by their evolutionary similarity to humans. Crystal Bioscience has expanded the
repertoire of transgenic animals by engineering HuMab chickens producing fully human sequence, high affinity mAbs. In addition to revealing novel epitopes and, therefore novel IP, the Crystal Platform yields mAbs recognizing murine orthologs
of human antigens that facilitate pre-clinical studies.
1:55 Session Break
2:10 Chairperson’s Remarks
Elizabeth England, Scientist, Antibody Discovery and Protein Engineering, MedImmune
2:15 Counteracting Tumor Evasion of Antibody Immunity by a Novel Therapeutic Strategy
Zhiqiang An, Ph.D., Professor and Robert A. Welch Distinguished University Chair in Chemistry, University of Texas
Health Science Center at Houston
Immune suppression is recognized as a hallmark of cancer and this notion is largely based on studies on cellular immunity. Our recent studies have demonstrated a potential new mechanism of cancer suppression of immunity by impairment of antibody
effector function mediated by proteolytic enzymes in the tumor microenvironment. Furthermore, we are exploring strategies to restore the lost functions to the damaged antibodies, which represent potentially new directions in cancer immunotherapy.
2:45 Discovering Antibodies to a Moving Target
Elizabeth England, Scientist, Antibody Discovery and Protein Engineering, MedImmune
MedImmune has shown that IL-33 forms disulphide bonds, resulting in large conformational changes. This occurs rapidly, posing a challenge in identifying antibodies that inhibit the action of IL-33. Through innovative use of mutant forms
of IL-33 and appropriate design of screening campaigns, a highly potent inhibitor of IL-33 was identified, a testament to how understanding of target structure and biology is key to the identification of potential therapeutic drug
candidates.
3:15 llamdA™ Next Generation VHH Library: Combining Immune Functionality with Synthetic Diversity
Guy Hermans, CSO, Isogenica Limited
We will present validation data from different selection campaigns, demonstrating our llamdA synthetic camelid domain antibody library consistently delivers a high number of binder with excellent affinity and broad epitope coverage, without
losing the beneficial solubility and stability properties as described for immune animals derived VHHs.
3:30 Improving Developability with Protein Surface Charge and Hydrophobic Patch Analysis
Nels Thorsteinson, Scientific Services Manager, Biologics, Chemical Computing Group
We describe a computational method for identifying and measuring hydrophobic and charged patches on the surface of the protein structure. An analysis of protein complexes in the PDB suggests that hydrophobic patches play an important
role in binding. Their application to reducing aggregation, improving solubility, and epitope mapping is demonstrated.
4:00 Refreshment Break in the Exhibit Hall with Poster Viewing
4:45 Problem-Solving Breakout Discussions
These interactive discussion groups are open to all attendees, speakers, sponsors, & exhibitors. Participants choose a specific breakout 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. Pre-registration to sign up for one of the topics will occur a week or two prior to the Event via the App.
Droplet Microfluidics for Antibody Discovery
Moderator: Christoph Merten, Ph.D., Group Leader Microfluidics, European Molecular Biology Laboratory (EMBL), Germany
- What kind of screens can be done on primary plasma cells from mice and humans?
- What are the specific advantages compared to other screening formats?
- How to get access to the technology?
5:45 Networking Reception in the Exhibit Hall with Poster Viewing
7:00 End of Day
THURSDAY, MAY 4
8:00 am Morning Coffee
8:30 Chairperson’s Remarks
Ruud de Wildt, Ph.D., Director, Head, Antibody Selections, Biopharm, GlaxoSmithKline
8:35 Modeling and Docking Antibody Structures with Rosetta
Jeliazko Jeliazkov, Research Assistant, Molecular Biophysics, Johns Hopkins University
Structures of antibodies in complex with their antigens can give insight into therapeutic mechanisms and suggest improved antibody designs. We have developed computational methods (1) to create atomically accurate models of antibodies
from their sequences and (2) to dock those models to antigens. This talk will present the methods, critically analyze their accuracy, and demonstrate applications to large-scale repertoire analysis as well as Celiac disease
and pulmonary hypertension.
9:05 Computational Design of de novo Anti-Influenza Minibinder Proteins
Christopher Pirie, Ph.D.,CEO, Virvio, Inc.
Rosetta computational design can create proteins that target neutralizing epitopes on Influenza hemagglutinin. These designed proteins bind like broadly neutralizing antibodies, but in a smaller molecule that is both more manufacturable
and stable. In animal models they inhibit the function of hemagglutinin and prevent viral infectivity. These computationally designed binders represent a new class of protein therapeutics for infectious diseases.
9:35 Computational Approaches in Antibody Design: Identifying and Reducing Liabilities Early in the Discovery Process
David Pearlman, Senior Principal Scientist, Schrödinger
Computational tools that can be used in the optimization process for putative antibody drug candidates have greatly improved in the past several years. Using the BioLuminate software platform, we describe both how these calculations
can be utilized for workflowed triage among multiple candidates, and how tools such as FEP can be used to suggest sequence engineering that can ameliorate identified liabilities such as aggregation propensity while maintaining
affinity and stability.
10:05 Coffee Break in the Exhibit Hall with Poster Viewing
11:05 Optimizing Antibodies for Targeting the Tau Protein for Alzheimer’s Disease and Other Tauopathies
James A. Ernst, Ph.D., Senior Scientist, gRED, Protein Chemistry and Neuroscience, Research
and Early Development, Genentech, Inc.
Tau is a prime therapeutic target for Alzheimer’s disease. We discovered that in pre-clinical models in vivo and in cell based experiments, effector function is not required
for targeting tau efficaciously with tau antibodies, and that attenuation of effector function could be advantageous. We propose that effector function status is an important consideration when designing therapeutic tau antibodies,
and potentially for antibodies against other neurodegeneration targets as well.
11:35 Progress and Challenges with the Isolation and Optimization of Antibodies against Multi-Spanning Transmembrane Targets
Ruud de Wildt, Ph.D., Director, Head, Antibody Selections, Biopharm, GlaxoSmithKline
Raising antibodies to complex cellular targets such as ion channels, G-protein coupled receptors (GPCRs) and other multi-spanning membrane targets is typically very challenging due to their complex nature and limited antigen availability.
This talk will describe the strategies implemented by GSK to successfully identify high potency neutralizing antibodies for such targets, using the ADIMABTM yeast-based platform and in vivo immunization approaches.
12:05 pm Targeting Islet Amyloid Polypeptide (IAPP) as an Immunotherapy for Type 2 Diabetes
Robin Barbour, Head, Antibody and Assay Development, Research, Prothena Biosciences
Islet Amyloid Polypeptide (IAPP) is a 37 amino acid aggregation prone peptide that is co-secreted with insulin, and the toxicity of IAPP aggregates are believed to contribute to the pathophysiology of type 2 diabetes. We used a
transgenic rat model that expresses human IAPP, which presents type 2 diabetes-relevant phenotypes such as pancreatic IAPP deposition and loss of insulin-secreting beta-cells, and showed that anti-IAPP immunotherapy slowed
the disease progression in this model.
12:35 End of Engineering Antibodies