Immediately following CHI's Phage Display Conference

The pipeline of therapeutic antibodies has been growing steadily, building on successful clinical trials, particularly in cancer. This growth will certainly accelerate, as genomic companies with many novel targets seek to employ antibodies to modulate these targets. In addition, studies employing antibodies, or antibody mimics, are being ramped up as part of the explosive growth of proteomics. All of these trends place a tremendous emphasis on the development of new approaches for faster antibody development, improved methods of selection and optimization, alternatives for production, and evaluation for novel applications. This conference will provide you with key updates on important developments in each of these areas.

Sponsoring Publications: 
Drug Discovery and Development
Human Antibodies
Immunotherapy Weekly
The Scientist
Protein Science

SCIENTIFIC ADVISORS
Dr. Andrew Bradbury, Los Alamos National Laboratories
Dr. Peter J. Hudson, CSIRO Health Sciences and Nutrition (Australia)
Dr. Hennie R. Hoogenboom, Dyax SA and University of Liège (Belgium)

ADDITIONAL SPEAKERS
Dr. Robert Balint, KaloBios, Inc.
Dr. Gerald Beste, Xerion Pharmaceuticals AG (Germany)
Dr. Ole H. Brekke, Affitech AS (Norway)
Dr. Antonino Cattaneo, University of Trieste (Italy)
Dr. Dominik Escher, ESBATech AG (Switzerland)
Dr. Lutz Jermutus, Cambridge Antibody Technology (UK)
Dr. Dev Kambhampati, Thermo Hybaid GmbH, (Germany)
Dr. Stephen Kingsmore, Molecular Staging Inc.
Dr. Sergey Kipriyanov, Affimed Therapeutics AG (Germany)
Dr. John M. Lambert, ImmunoGen, Inc.
Dr. James Marks, University of California, San Francisco
Dr. Jennie Mather, Raven Biotechnologies
Dr. Serge Muyldermans, Vrije Universiteit Brussel (Belgium)
Dr. Stewart Nuttall, CSIRO Health Sciences and Nutrition (Australia)
Dr. Peter Pavlik, Los Alamos National Laboratories
Dr. Christoph Rader, The Scripps Research Institute
Dr. Arvind Rajpal, University of California, Berkeley
Dr. Daniele Sblattero, University of Trieste (Italy)
Prof. Dr. Arne Skerra, Pieris ProteoLab AG (Germany)
Dr. Eskil Söderlind, BioInvent Therapeutic AB (Sweden)
Dr. Boris Steipe, University of Toronto (Canada)
Dr. Alain Tissot, Cytos Biotechnology AG (Switzerland)
Dr. Sydney Welt, Memorial Sloan Kettering Cancer Center
Dr. Li Zhu, Genetastix Corporation

SELECTION
Streamlined Display Platform for the Fast Isolation and Affinity Maturation of Human Antibodies
Application of the n-CoDeR™ Antibody Libraries
In Vitro Selection and Evolution in Antibody Drug Discovery
Antibody Engineering with TACZYMEs (Target-Activated Enzymes)

IMPROVEMENTS AND ANTIBODY ALTERNATIVES
From Butterflies to ANTICALINS: Small Engineered Ligand-Binding Proteins as a Better Antibody Alternative
Current Status of Single Domain Camel Antibodies
Pepbodies™ - Novel Antibody Fragments with Natural Immunoglobulin Effector Functions
Single Domain Binding Reagents from Shark NAR Antibodies

INTRACELLULAR ANTIBODIES
IACT: The Intracellular Antibody Capture Technology
Highly Efficient Yeast Two-Hybrid System-Based Screening and Affinity Maturation System for Human Monoclonal Antibodies
The Canonical Sequence Approximation: A Rational Route to Intrabodies
Selecting the Right Antibodies for Intracellular Applications
Intracellular Stability Determines Efficacy of Intrabodies

THERAPEUTIC APPLICATIONS
A Cell-Based Approach to the Simultaneous Discovery of Targets and Therapeutic Antibodies
Lessons from Potent Neurotoxin Neutralization by Phage Antibodies
Novel Recombinant Bispecific Molecules for Immunotherapy of Blood Malignancies
Immunodrugs™: A New Class of Biopharmaceuticals to Treat Chronic Diseases
Preliminary Report of a Phase I Study of Combination Chemotherapy and Humanized A33 (huA33) Immunotherapy
Targeting Tumor Angiogenesis with Recombinant Antibodies
Conferring Therapeutic Potency to Anticancer Antibodies through Conjugation to Maytansinoid Drugs

RESEARCH AND DRUG DEVELOPMENT APPLICATIONS
Applying Phage Antibodies to Functional Genomics
Recombinant Antibodies and Chromophore-Assisted Laser Inactivation (CALI) for Functional Protein Knockout
RCA-Enabled Antibody Arrays for Biomarker Identification in Serum
What Patient Phage Antibody Libraries Can Tell Us about Autoimmunity
Investigating Antigen-Antibody Interactions Using a Highly Specific Protein Chip Platform

TUESDAY, APRIL 23

6:00-7:00pm Early Registration, Poster and Exhibit Setup

 

WEDNESDAY, APRIL 24

7:30 Registration, Poster and Exhibit Viewing, and Light Continental Breakfast

 

Selection

8:30 Welcome by Session Chairperson
Dr. Andrew Bradbury, Los Alamos National Laboratories

8:40 A Streamlined Display Platform for the Fast Isolation and Affinity Maturation of Human Antibodies
Dr. Hennie R. Hoogenboom, Senior Vice President, Discovery, Dyax Corp., and Professor, University of Liège
We describe an integrated technology platform with novel cloning, display and selection methods, that is used to develop human antibodies. Core to our platform is a novel cloning method which is based on site-specific cleavage of ss-DNA. Examples will be given of the utility of this procedure for the construction of human Fab libraries and, in a further novel application, for the site-directed diversification of antibody genes. Complemented by automated phage display selection systems, and yeast-display based affinity selection procedures, this platform provides rapid access to high affinity human antibody biopharmaceuticals.

9:10 Application of the n-CoDeR™ Antibody Libraries
Dr. Eskil Söderlind, Associate Professor, Director Scientific Liaisons, BioInvent Therapeutic AB
The n-CoDeR antibody libraries are built on a single master framework into which diverse in vivo formed complementarity determining regions (CDRs) are allowed to recombine. These CDRs are sampled from in vivo processed and proofread gene sequences, thus ensuring an optimal level of correctly folded and reactive molecules. Highly specific and functional antibody fragments have been selected using the n-CoDeR libraries and the latest results will be presented.

9:40 In Vitro Selection and Evolution in Antibody Drug Discovery
Dr. Lutz Jermutus, Display Technology Development, Cambridge Antibody Technology
Ribosome display is a powerful in vitro selection system that can be utilized in both the primary isolation and the evolution of antibody drug candidates. Data will be presented to illustrate how the technique has been used to generate cytokine-neutralizing antibodies. Primary selection strategies, directed evolution strategies, high-throughput screening data and results of cell-based assays demonstrating the biological activity of the in vitro selected antibodies will be explained. These examples will be used to highlight the various specific advantages of ribosome display as compared to other selection methods such as phage display or surface display. (Coauthors: Maria Groves, Ross Stewart, George Thom, Emma de Vries, Svava Wetzel)

10:10 Antibody Engineering with TACZYMEs (Target-Activated Enzymes)
Dr. Robert Balint, Founder and Chief Technology Officer, KaloBios, Inc.
We will describe the development of enzyme sensors (TACZYMES) which can be activated or inactivated by antigen-antibody interactions, and the use of these sensors in cell-based systems for epitope-guided selection and affinity maturation of human antibodies for therapeutic applications.

10:40 Poster and Exhibit Viewing, Refreshment Break

 

Improvements and Antibody Alternatives

11:15 Comments by Session Chairperson
Dr. Hennie R. Hoogenboom

11:20 From Butterflies to ANTICALINS: Small Engineered Ligand-Binding Proteins as a Better Antibody Alternative
Dr. Arne Skerra, Co-Founder and Chairman, Pieris ProteoLab AG
ANTICALINS are a novel class of soluble receptor proteins based on the lipocalin scaffold. Their robust beta-barrel architecture supports four loops, which form a ligand pocket and which can be extensively reshaped to specifically bind small molecules as well as large proteins. Starting with a lipocalin from the butterfly Pieris brassicae, human lipocalin structures have recently been recruited in order to construct ANTICALINS as target recognition modules for therapeutic application.

11:50 Current Status of Single Domain Camel Antibodies
Dr. Serge Muyldermans, Department "Ultrastructure", Vlaams Interuniversitair Instituut voor Biotechnologie, Vrije Universiteit Brussel
Camelids possess functional heavy-chain antibodies that lack light chains. These antibodies are assembled from dedicated V and Cgamma genes. After immunizing dromedaries or llamas and adapting the phage display technology we isolate antigen-specific single-domain antibody fragments composed of variable domains of HCAbs (i.e., VHH). The antigen-binding properties (affinity, X-ray structure) of VHHs were analyzed, and several applications are envisaged where the single-domain nature, the intrinsic high stability and the potential to reach hidden epitopes, of these minimal-sized antigen binders offer particular advantages.

12:20 Lunch (on your own)

1:35 Comments by Session Chairperson
Dr. Hennie R. Hoogenboom

1:40 Pepbodies™: Novel Antibody Fragments with Natural Immunoglobulin Effector Functions
Dr. Ole H. Brekke, Director, Explorative Research, Affitech AS
Antibodies bind antigens and eliminate them via immunoglobulin (Ig) effector functions such as, activation of the complement system and interaction with cellular receptors. All effector functions are associated with the immunoglobulin Fc-region. Fully functional immunoglobulins must be glycosylated and thus be produced in eukaryotic expression systems. Fv, scFv, or Fab fragments can be successfully expressed in E. coli, but these compounds lack the Fc-region and thus also the effector functions. We have developed small bacterially produced antibody fragments that can cross link antigen and antibody effector molecules. We have termed these molecules Pepbodies™. Pepbodies™ are binding molecules comprising antigen-binding sites genetically fused to peptides that display one or more of the effector functions associated with the Fc-region. (Coauthors: Vigdis Lauvrak and Inger Sandlie, Division of Molecular Cell Biology, Department of Biology, University of Oslo)

2:10 Single Domain Binding Reagents from Shark NAR Antibodies
Dr. Stewart Nuttall, CSIRO Health Sciences and Nutrition, and CRC for Diagnostics
Small domains provide an important alternative scaffold to complement existing peptide and antibody libraries. The New Antigen Receptor (NAR) from sharks encapsulates binding affinity in a single immunoglobulin variable-like domain, and is a candidate for the minimal antibody-based antigen-binding fragment as it utilizes only CDR1 and -3 loops. We have engineered these single domain fragments for expression in E. coli, and produced in vitro bacteriophage displayed libraries by CDR mutagenesis, mimicking structural features such as inter-CDR disulphide linkages found in the wild-type antibodies. The results of library panning experiments against a range of antigens will be presented, including data on protein expression, affinities, and initial structural determinations.

2:40 Panel Discussion with Questions and Answers from Audience for All Session Speakers

Intracellular Antibodies

3:00 Comments by Session Chairperson
Dr. Dominik Escher, Chief Executive Officer, ESBATech AG

3:05 IACT: The Intracellular Antibody Capture Technology
Dr. Antonino Cattaneo, Professor of Biophysics at SISSA, International School for Advanced Studies;President of Lay Line Genomics SpA
The selection of functional intracellular antibodies by the intracellular antibody capture technology will be described. Antibodies against a number of different intracellular antigens were selected de novo and used for protein knock out in mammalian cell lines and primary neurons. Sequences of antibodies selected with IACT were compiled in a database of validated intrabody sequences, and used to establish a consensus sequence for functional intracellular antibody frameworks.

3:35 Highly Efficient Yeast Two-Hybrid System-Based Screening and Affinity Maturation System for Human Monoclonal Antibodies
Dr. Li Zhu, President and Chief Executive Officer, Genetastix Corporation
Genetastix has developed a highly efficient and versatile antibody screening and affinity maturation system based on yeast two-hybrid system. This system allows us to isolate human scFv antibodies with sub-nanomolar level of affinity within a very short time. We have used this system to generate antibodies against disease-related antigens that are normally expressed intracellularly, secreted, or membrane-bound.

4:05 Poster and Exhibit Viewing, Refreshment Break

4:45 The Canonical Sequence Approximation: A Rational Route to Intrabodies
Dr. Boris Steipe, Associate Professor, Department of Biochemistry, Program in Proteomics and Bioinformatics University of Toronto
To address the stability problems of immunoglobulin domains in the reducing environment of the cytoplasm, we have devised a strategy for rational stability engineering based on consensus sequences. Point mutations are predicted from sequence alignments and provide incremental increases of stability. These can be combined in existing domains, or used for the design of hyperstable frameworks and routinely allow expression of the target domains as soluble intrabodies with high yield. The method is straightforward, does not require knowledge of the structure, is applicable to scFvs as well as isolated domains and improves expression yields and solubility at the same time.

5:15 Selecting the Right Antibodies for Intracellular Applications
Dr. Dominik Escher
Applications of single-chain antibodies within the cell have a great promise, both in target validation and therapeutics. Unfortunately the intracellular environment differs from the natural secretion pathway of antibodies. Within the cell, the reducing environment prevents formation of disulfide bonds which are known to be important for the stability, the correct folding and therefore the function of the majority of single-chain antibodies. Thus most of the tested single-chain antibodies have a reduced affinity to their target or are even completely not functional within the cell. We have developed a novel in vivo technology which identifies single-chain antibodies suitable for intracellular applications. This selection of optimal frameworks is carried out independently of any antigen interaction. Results on selected frameworks will be presented.

5:45 Intracellular Stability Determines Efficacy of Intrabodies
Dr. Arvind Rajpal, Department of MCB, Division of Immunology, University of California, Berkeley
We generated a panel of five scFvs, directed against catalytically inactive murine caspase-3, by screening phage displayed libraries, in our efforts to determine the crucial principles that dictate the efficiency of retargeting antigen intracellularly. Our results demonstrate that the intrinsic stability, rather than the affinity for the antigen, plays a crucial role in intracellular efficacy of the intrabody.

6:15 Panel Discussion with Questions and Answers from Audience for All Session Speakers

6:30-7:45 Networking Reception (sponsored by Cambridge Healthtech Institute)

 

THURSDAY, APRIL 25

8:00am Poster and Exhibit Viewing and Light Continental Breakfast

Therapeutic Applications

8:30 Comments by Session Chairperson
Dr. Dev Kambhampati, Product Manager, XNA on Gold Microarrays, Interactiva Division, Thermo Hybaid GmbH

8:35 A Cell-Based Approach to the Simultaneous Discovery of Targets and Therapeutic Antibodies for the Treatment of Cancer
Dr. Jennie Mather, President and Founder, Raven Biotechnologies
Raven Biotechnologies' proprietary, approach to drug discovery uses tissue-specific progenitor cell lines and proprietary methods optimized for producing monoclonal antibodies (Mabs) targeting cell surface proteins. This integrated approach rapidly produces targets, including targets not available through genomics approaches, and therapeutic candidates that are comprehensively described in a functional context. Raven is initially directing its technology toward the discovery of monoclonal antibody therapeutics for cancer.

9:05 Lessons from Potent Neurotoxin Neutralization by Phage Antibodies
Dr. James Marks, Professor, Anesthesia & Pharmaceutical Chemistry, University of California, San Francisco
The botulinum neurotoxins (BoNTs) cause the paralytic human disease botulism and are also one of the highest-risk threat agents for bioterrorism. We have utilized immune phage antibody libraries constructed from humans, mice, and mice transgenic for the human Ig locus to dissect the immune response to toxin and to identify the requirements for potent toxin neutralization. Results will be presented showing a general route to achieving highly potent toxin neutralization applicable not only to BoNTs, but to other biothreat agents as well. Such antibodies can be used for treatment and prevention of botulism resulting from natural exposure or bioterrorism.

9:35 Novel Recombinant Bispecific Molecules for Immunotherapy of Blood Malignancies
Dr. Sergey Kipriyanov, Project Leader, Affimed Therapeutics AG
Recombinant antibody technology provides powerful tools for enhancing the potency of therapeutic bispecific antibodies. The presentation will describe construction and pre-clinical examination of recombinant molecules retargeting different populations of human effector cells towards the non-Hodgkin's lymphoma cells. (Coauthors: Björn Cochlovius, Holger Schäfer, Fabrice Le Gall, Uwe Reusch, Gerhard Moldenhauer, and Melvyn Little)

10:05 Immunodrugs™: A New Class of Biopharmaceuticals to Treat Chronic Diseases
Dr. Alain Tissot, Senior Scientist, Cytos Biotechnology AG
The Immunodrugs are designed to instruct the patient's immune system to produce a desired therapeutic antibody that blocks or activates a disease-associated antigen with the goal of prevention, long-lasting stabilization or reversion of the disease process. This is achieved by presentation of the selected disease-related protein to the patient's immune system in a highly repetitive array known to efficiently trigger an immune response.

10:35 Poster and Exhibit Viewing, Refreshment Break

11:05 A Preliminary Report of a Phase I Study of Combination Chemotherapy and Humanized A33 (huA33) Immunotherapy in Patients with Advanced Colorectal Cancer
Dr. Sydney Welt, Memorial Sloan-Kettering Cancer Center
Humanized A33 IgG1 antibody (huA33) has modest anti-tumor activity when used as a single agent. However, when chemoresistant colorectal cancer patients were treated with chemotherapy after a huA33 therapy protocol, major responses were observed in a significant portion of patients. This observation led to the present study, which was designed to determine if huA33 immunotherapy and chemotherapy can be combined and if major responses are induced. The current report indicates that huA33 immunotherapy can be combined with chemotherapy without additional toxicities and that major durable responses are obtained in a substantial portion of the patients.

11:35 Targeting Tumor Angiogenesis with Recombinant Antibodies
Dr. Christoph Rader, Department of Molecular Biology, The Scripps Research Institute
Using phage display, we have generated, humanized, and evolved antibodies to defined molecular targets that are functionally implicated in tumor angiogenesis. Among the selected features were very high affinity and cross-reactivity between mouse and human antigen. Based on novel vectors for whole antibody expression and adenoviral delivery of intrabodies, downstream constructs of the selected antibodies were engineered and evaluated for antiangiogenic therapy in mouse models of human cancer.

12:05 Tumor-Activated Prodrugs: Conferring Therapeutic Potency to Anticancer Antibodies Through Conjugation to Maytansinoid Drugs
Dr. John M. Lambert, Senior Vice President, Pharmaceutical Development, ImmunoGen, Inc.
Monoclonal antibodies often show only modest antitumor activity as single agents. A new generation of antibody-drug conjugates containing maytansinoids act like Tumor-Activated Prodrugs (TAPs). Such TAPs display potent antitumor activity in tumor xenografts models at nontoxic doses and are now undergoing clinical evaluation in Phase I trials.

12:35 Panel Discussion with Questions & Answers from Audience for All Session Speakers

12:55 Luncheon (sponsored by Cambridge Healthtech Institute)

Research and Drug Development Applications

2:15 Comments by Session Chairperson
Dr. Dev Kambhampati

2:20 Applying Phage Antibodies to Functional Genomics
Dr. Peter Pavlik, Los Alamos National Laboratories
While selection of phage antibodies from large libraries is relatively trivial, their use in downstream proteomics applications can be complicated by poor expression levels and monomeric affinities. In this talk approaches to overcome these problems and automate selection will be discussed.

2:50 Use of Recombinant Antibodies and Chromophore-Assisted Laser Inactivation (CALI) for Functional Protein Knockout
Dr. Gerald Beste, Director of Combinatorial Technology, Xerion Pharmaceuticals AG
Xerion employs its functional proteomics technology for the systematic functional analysis and validation of proteins in cell-based assays. The centerpiece of the technology platform is based on Chromophore Assisted Laser Inactivation (CALI), which provides a timely and locally restricted protein knockout. In contrast to "genetic" knockouts or antisense technologies this target validation approach does not suffer from problems with gene compensation and the low correlation between mRNA and protein expression.

3:45 RCA-Enabled Antibody Arrays for Biomarker Identification in Serum
Dr. Stephen Kingsmore, Molecular Staging Inc.
A significant component of the emerging discipline of proteomics will be measurement of biologically relevant groups of proteins in many disease states and treatments. Conventional cognate protein measurement methods lack the sensitivity, scalability, and reproducibility needed for simultaneous quantitation of many proteins in parallel. Attachment of an oligonucleotide to an anti-biotin antibody enables the use of Rolling Circle Amplification (RCA) as a generic signal amplification method for protein-ligand interactions on microarrays. We have constructed such a microarray for measurement of 75 cytokines, chemokines and growth factors by use of specific antibody pairs and signal amplification by RCA. Using this chip, we have examined the time course of cytokine secretion by human dendritic cells cultured under different conditions, measured changes in protein levels in serum from SCID patients who have undergone bone marrow transplant, and identified markers in cord blood from neonates who go on to develop cerebral palsy. Work is in progress to develop a second-generation chip for measurement of drug toxicity markers.

3:20 Poster and Exhibit Viewing, Refreshment Break

4:15 What Patient Phage Antibody Libraries Can Tell Us about Autoimmunity: The Case of Celiac Disease
Dr. Daniele Sblattero, Department of Biology, University of Trieste (Italy)
Although autoantibodies are characteristic of many autoimmune disease, their involvement in the pathological process has been debated. We have made phage antibody libraries from lymphocyte subsets of celiac disease patients. These libraries have revealed to be a reliable source of autoantibodies used in a variety of biological assays. These experiments suggest that celiac disease autoantibodies have indeed an important role in the pathogenesis of the mucosal lesion typical of celiac disease.

4:45 Investigating Antigen-Antibody Interactions Using a Highly Specific Protein Chip Platform: XNA on Gold™ Microarrays
Dr. Dev Kambhampati
We report a novel microarray platform, XNA on Gold, for screening proteins and oligosaccharides. The XNA protein and glycochip can be used to screen antigen-antibody interactions with high specificity, generating data with high signal-to-noise ratios. Different detection modes for monitoring recombinant antibodies and protein interactions will be discussed along with the current challenges in developing novel diagnostic kits.

5:15 Panel Discussion with Questions and Answers from Audience with All Session Speakers

5:30 Close of Conference

CALL FOR POSTERS
Cambridge Healthtech Institute encourages attendees to gain further exposure by presenting their work in the poster sessions. Please fill out the registration form, with the poster title and primary author. To ensure inclusion in the conference binder, a one-page summary must be submitted and registration must be paid in full by March 22, 2002.  Click here for poster instructions

CALL FOR SPONSORS AND EXHIBITORS
Recombinant antibody technology has a wide variety of uses for numerous types of scientific researchers. Most recently, the emphasis has been towards cancer research and treatments and to their applications in the exploding field of proteomics. The role of phage display in antibody production will be an integral part of this meeting, thus creating a perfect conclusion to the Phage Display conference held immediately prior. We strongly encourage any company with services or products related to antibody production, selection and modification, immunogenicity prediction, intracellular antibodies, antibody libraries, peptide arrays, antibody alternatives, phage display, and disease research to consider sponsoring or exhibiting at this event. Certainly sponsorship is the best way to prominently elevate your company's presence and influence at this conference. Although exhibit space can be purchased separately, many sponsorship packages will include a booth. The early deadline of January 4, 2002 is fast approaching-registering to exhibit before that date will save your company up to $600! For more information on sponsorship opportunities or to reserve a booth, please contact Jim MacNeil at 781-972-5441 or jmacneil@healthtech.com.

Phage Display Technologies Exhibitors:
The following companies are exhibiting as of April 20, 2002
Dyax Corporation
TSI Incorporated

Recombinant Antibodies Exhibitors:
The following companies are exhibiting as of April 20, 2002
Altus Biologics, Inc.
TSI Incorporated
Sapidyne Instruments Inc.

HOTEL INFORMATION
University Park Hotel @ MIT
20 Sidney Street
Cambridge, MA 02139
T: 617-577-0200 o 800-222-8733
F: 617-494-8366
W: www.univparkhotel.com
Room Rates: $195/S o $205/D
Cut-off-Date: April 1, 2002
Please call the hotel directly to make your room reservation. Identify yourself as a Cambridge Healthtech Institute conference attendee to receive the reduced room rate. Reservations made after the cut-off date or after the group room block has been filled (whichever comes first) will be accepted on a space-and-rate-availability basis. Rooms are limited, so please book early.

TRAVEL INFORMATION
Special Airline Discounts Available
Special zone and discount fares have been established on United Airlines for this conference. Please call United Airlines Meeting Reservations Center directly at 800-521-4041. You must reference ID #579YS.