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. Engineering Antibodies presents the state of the art in technologies used by protein engineers working at the discovery and design stage to quickly and efficiently craft biotherapeutics directed at the most elusive targets and biological functions.
WEDNESDAY, APRIL 27
7:00 am Registration and Morning Coffee
8:00 Chairperson’s Remarks
James Ernst, Ph.D., Senior Scientist, Therapeutic Proteins, Protein Chemistry, Genentech
8:10 KEYNOTE PRESENTATION:
From Synthetic Antibodies to Synthetic Proteins
Sachdev Sidhu, Ph.D., Professor, Molecular Genetics, University of Toronto
We have used structure-based combinatorial design to build simple yet highly functional synthetic antibody libraries. The principles learned from antibody design have been applied to alternative natural scaffolds including ubiquitin and SH2 domains. Finally, we have developed highly optimized small scaffolds that are amenable to chemical synthesis to build D-amino acid binding proteins with traits favorable for therapeutic applications.
8:40 Retargeting T Cells in Hematologic Malignancies
Gerhard Zugmaier, M.D., Professor, Internal Medicine, Hematology/Oncology, Marburg University; Executive Medical Director, Global Development, Amgen Research Munich
Blinatumomab, the most advanced bispecific T cell engager (BiTE), targets CD19 positive B cells. In patients with minimal residual disease (MRD) of B-precursor Acute Lymphoblastic Leukemia (ALL) blinatumomab induced molecular remissions resulting in an estimated relapse-free survival (RFS) of 60% at a follow-up of 31 months. In patients with relapsed/refractory B-precursor ALL blinatumomab induced remissions enabling successful allogeneic hematopoietic stem cell transplantation (HSCT). Blinatumomab induced durable responses in Non-Hodgkin Lymphoma.
9:10 Engineering Molecules for Cellular Uptake
Wouter Verdurmen, Ph.D., Postdoctoral Fellow, Biochemistry, University of Zurich
To accurately measure the delivery of proteins to the cytosol, the biotin ligase assay was developed that allows the objective quantification of cytosolic delivery. The method has been employed for comparing the cytosolic delivery of various engineered protein translocation mechanisms. This has enabled the identification of mechanisms that efficiently transport protein cargoes to the cytosol, as well as generated a detailed understanding of the characteristics that underlie efficient transport.
9:40 Blood-Brain Barrier Penetrating Dual Specific Binding Proteins for Treating Brain and Neurological Diseases
Denise Karaoglu Hanzatian, Ph.D., Principal Research Scientist, AbbVie Bioresearch Center
While naturally protective, the blood–brain barrier (BBB) provides a challenge for drug development as most of the small organic molecule drugs and nearly all biologics do not cross the BBB to reach therapeutically relevant concentrations. Here, we present the generation and expression of DVD-Ig proteins capable of binding specific disease targets in the brain. Uptake, retention and the elevated functional activity of DVD-Ig proteins in brain will be demonstrated.
10:10 Coffee Break in the Exhibit Hall with Poster Viewing
10:55 An Fc Engineering Approach that Modulates Antibody-Dependent Cytokine Release Without Altering Cell-Killing Functions
Michelle Kinder, Ph.D., Scientist Immuno-Oncology Discovery, Janssen Research & Development, LLC
In addition to Fc-mediated target cell-killing, therapeutic antibodies can elicit cytokine release from peripheral blood mononuclear cells and macrophages that can influence disease microenvironments and therapeutic outcomes. We describe an Fc engineering approach that results in macrophage-mediated phagocytosis and subsequent cell-killing without eliciting cytokine release from macrophages. When peripheral blood mononuclear cells are used as effectors, the resulting variants have similar cell-killing and cytokine release compared to IgG1.
11:25 Functional Antibody Screening Technology for Challenging Targets
Byeong Doo Song, Ph.D., President, Scripps Korea Antibody Institute
Current antibody screening technology is limited mostly to simple targets and has not been applied successfully to complex targets such as GPCRs, or ion channels. Scripps Korea Antibody Institute (SKAI) has developed functional antibody screening technology (FAST) that enables discovery of functional antibodies for targets of any structural complexity through individual & functional screening of spatially addressed antibody library using cell-based assays. Functional antibodies for GPCRs will be discussed.
11:55 In vitro Fab Display: A Next-Generation Antibody Discovery Platform
Ryan Stafford, Ph.D., Associate Director, Protein Engineering, Sutro Biopharma, Inc.
Once considered unviable, we have developed robust methods for discovering antibodies from large Fab libraries using ribosome display. Selected Fabs can be reformatted into full-length antibodies and directly expressed as IgGs in our cell-free platform for rapid screening. Lead IgGs usually have high affinity and excellent biophysical properties, but can also be easily affinity matured if necessary. Use of a common LC enables generation of IgG bispecifics.
12:25 pm Analysis of the Genomics and Transcriptomics of CHO-K1 to Accelerate and De-risk Biologics Development
Pierre-Alain Girod, Ph.D., CSO, Selexis, SA
Detailed genomic and transcriptomic analysis of CHO-K1 can provide comprehensive data to support cell line engineering and genomic characterization of research cell banks (RCB). We will present our CHO-K1 genome and transcriptome data and its use to enable the generation of RCBs for difficult-to-express proteins as well as support regulatory packages.
12:55 Luncheon Presentation I: Precisely Controlled, Highly Diverse
Gene Mutant Libraries for Bioherapeutic Discovery and Development
Emily Leproust, Ph.D., CEO, Twist Bioscience
Critical to the success of bio-therapeutic discovery and development process is the availability of high quality libraries. By implementing a rational design approach and combining this with our expertise in synthetic DNA, Twist Bioscience delivers inexpensive, high diversity, high precision libraries with reduced turn-around time enabling optimization and acceleration of the design, build and test cycle.
1:25 Luncheon Presentation II: Uncovering Receptor Targets and Off-Targets Using Cell Microarray Technology
Jim Freeth, Ph.D., Managing Director, Retrogenix Ltd.
Human cell microarray screening enables rapid discovery of the primary cell surface receptors and off-targets of antibodies, proteins, viruses and small molecules. Case studies from pharmaceutical partners will demonstrate how this unrivalled platform has been used to: 1) Uncover novel targets from antibody phenotypic screening approaches; 2) Identify receptors for protein ligands in normal and disease processes, such as immune checkpoint interactions, and 3) Screen for unintended off-target activities of biotherapeutics.
1:55 Session Break
2:10 Chairperson’s Remarks
Carolyn Cuff, Ph.D., Leader, Translational Research & Investigation Unit, AbbVie Bioresearch Center, Inc.
2:15 A Novel Tissue-Specific Agonist of the FGF21 Pathway for the Treatment of Type 2 Diabetes
James Ernst, Ph.D., Senior Scientist, Therapeutic Proteins, Protein Chemistry, Genentech
Activation of the FGF21 pathway has been shown to improve several features of type 2 diabetes in mice and humans. We have discovered a novel bispecific antibody that mimics the function and metabolic effects of FGF21. Treatment with this antibody improves glycemic and lipid profiles in mouse disease models. This talk will describe the discovery of an antibody that binds Klotho-Beta and FGFR1, thereby stimulating the cognate FGF21 co-receptor complex in adipose tissues.
2:45 Are PCSK9 Inhibitors the Next Breakthrough in the Cardiovascular Field?
Robert P. Giugliano, M.D., Senior Investigator, TIMI Study Group, Staff Physician, Cardiovascular Medicine, Brigham and Women’s Hospital; Associate Professor, Medicine, Harvard Medical School
A potent new class of monoclonal antibodies directed again the PCSK9 protein represents the newest and most potent LDL-cholesterol lowering drugs currently available for clinical use. This talk will review their development, clinical efficacy and safety data, and the large phase III studies that are ongoing. The role of PCSK9 inhibition in the future management of patients with hypercholesterolemia will be explored.
3:15 Creating Focused Mutant Libraries for Protein Engineering
Nels Thorsteinson, Scientific Services Manager, Biologics, Chemical Computing Group
Protein engineering plays a pivotal role in modulating the function, activity and physical properties of biologics. However, the efficient search and evaluation of an excessively large sequence design space is challenging. 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.
3:45 Refreshment Break in the Exhibit Hall with Poster Viewing
4:45 Problem-Solving Breakout Discussions
Developing Technologies for Antibody Discovery and Engineering
James Ernst, Ph.D., Senior Scientist, Therapeutic Proteins, Protein Chemistry, Genentech, Inc.
- Antigen preparation for immunization
- Novel receptor targeting and activation
- Receptor clustering and internalization
- Predicting and addressing atypical pharmacokinetics
Next Generation Sequencing in Antibody Discovery and Engineering
Sai Reddy, Assistant Professor, ETH Zurich, Switzerland
- NGS methods for analyzing antibody repertoires
- Applications of NGS data for antibody engineering and discovery
- Challenges in data analysis and interpretation of NGS data
- Advanced tools for data correction
Use of Structural Biology Tools to Support Antibody Discovery
Jian Payandeh, Ph.D., Scientist, Structural Biology, Genentech, Inc.
- What are common challenges in your antibody discovery campaigns?
- What biophysical methods do you employ to characterize your Abs and antigens? Which are the most essential or powerful?
- Where are potential opportunities or synergies between different experimental approaches, and how are you taking advantage of these?
- When (and how) do you employ the use of surrogate antigens? What works well, what doesn’t?
5:45 Networking Reception in the Exhibit Hall with Poster Viewing
7:00 End of Day
THURSDAY, APRIL 28
8:00 am Morning Coffee
8:30 Chairperson’s Remarks
Sai Reddy, Ph.D., Assistant Professor, Biosystems Science and Engineering, ETH, Zurich, Switzerland
8:35 High-Throughput Conformational Epitope Mapping to Guide Design of Structure-Based Vaccines
Timothy Whitehead, Ph.D., Assistant Professor, Chemical Engineering and Materials Science, Michigan State University
We have developed a novel method for rapid determination of fine conformational epitopes. This platform technology involves deep sequencing of yeast displayed antigen libraries. I will present determination of critical (and previously unknown) neutralizing epitopes for pertussis toxin and a breast cancer target. Further, I will discuss methodological improvements in throughput and cost needed to integrate epitope mapping upstream in candidate selection. Implications for structural vaccine design will be discussed.
9:05 Designer Libraries for Soluble, Membrane Anchored and Membrane Spanning Proteases for Monitoring Their Role in Disease
Charles S. Craik, Ph.D., Professor, Pharmaceutical Chemistry, University of California, San Francisco
Selective tools for studying an individual protease are important for characterizing its function and understanding its role in disease. Targeting specific proteases is challenging due to the high structural homology of enzymes in a given family. Our results highlight the value of biased Fab libraries and structure-guided design for efficient identification of protease inhibitors and development of selective antibody-based tools for determining the structure and function of serine proteases in disease and normal physiology.
9:35 Computational Advances in Antibody Design: Toward Improved
Optimization and Selection
David Pearlman, Ph.D., Senior Principal Scientist, Schrödinger
Recent computational advances hold significant promise both for improved prediction of antibody structure from sequence, and for the ability to precisely calculate physically relevant properties such as affinity and stability. When combined with additional theoretical approaches to identify liabilities, we can use these tools to variously optimize a lead antibody candidate and triage among multiple potential leads.
10:05 Coffee Break in the Exhibit Hall with Poster Viewing
11:05 Unleashing the Structural Biology Toolbox to Facilitate Antibody Discovery
Jian Payandeh, Ph.D., Scientist, Structural Biology, Genentech
Structural biology can be leveraged in many ways to support therapeutic antibody discovery. Traditional applications at Genentech include facilitating structure-based antibody engineering efforts and understanding the mechanisms of antibody action. We have used insights from structural biology to engineer antigens and help guide our discovery campaigns, and we have implemented a high-throughput screening platform that streamlines the generation of challenging antigens, including ion channels, receptors, and GPCRs. Applications from our structural biology efforts will be described.
11:35 Human Antibody Transgenic Rabbits
Alain C. Tissot, Ph.D., Head, Immune Biology, Large Molecule Research, Roche Pharma Research & Early Development, Roche Innovation Center Penzberg, Roche Diagnostics GmbH, Switzerland
Antibodies generated in animal hosts represent the larger part of marketed therapeutic antibodies. Their generation undergoes positive and negative selection, delivering antibodies with robust therapeutic properties. Rabbits use a distinct diversity generation mechanism, which has long been exploited to achieve very high specificity. Humanization is, however, subsequently necessary. We report here on the generation of rabbits transgenic for human immunoglobulin genes, yielding therapeutic candidates of high affinity and specificity.
12:05 pm ABT-122, an Anti-TNF/IL-17 Dual Variable Domain Immunoglobulin (DVD-IgTM), Mechanisms of Translation: Bench to Bedside and Back Again
Carolyn Cuff, Ph.D., Leader, Translational Research & Investigation Unit, AbbVie Bioresearch Center, Inc.
ABT-122 is an anti-TNF/IL-17 dual variable domain immunoglobulin (DVD-Ig™) currently in Phase II clinical trials for Rheumatoid Arthritis (RA) and Psoriatic Arthritis (PsA). As these chemokine receptors and cytokine responses have been suggested to play a role in disease pathology or its resolution, these data suggest that dual blockade of TNF and IL-17 by ABT-122 could provide a new therapeutic approach to patients with RA and immune mediated inflammatory diseases.
12:35 End of Engineering Antibodies
5:15 Registration for Dinner Short Courses*