Monoclonal antibodies are quickly becoming key resources in the therapeutic, diagnostic and drug discovery fields. With the phenomenal growth in the market for monoclonal antibodies, it becomes even more critical to explore diverse methods of design, production and application of technologies to provide the antibodies needed.  This conference will provide researchers with key updates on important developments in the monoclonal antibody field covering generation, optimization, drug discovery research, and in vivo diagnostics, as well as case studies of successful mAb science.

THURSDAY APRIL, 9

12:00pm Conference Registration

Advances in Basic Science

1:30         Chairperson’s Opening Remarks

1:40         Developmental Trends in Antibody Fragment Therapeutics

Aaron Nelson, Ph.D., Tufts University School of Medicine

Monoclonal antibody derived fragments are a nascent category of targeted therapeutic agents, and a seemingly endless array of molecules can be engineered from various antibody domains.  We sought to determine trends in the development of 38 preclinical antibody fragments and 47 therapeutic molecules that have entered clinical study sponsored by commercial firms.  Our findings indicate antibody fragment therapy is an emerging technology that faces a major hurdle at the proof-of-clinical-concept stage.  Development may shift from FABs to scFvs and third-generation molecules, and from oncology indications to immunologic diseases.

2:10         Extending the Plasma Half-life of Antibody Fragments and Other Biopharmaceutical Proteins via Genetic Fusion with Homo-amino-acid Polymer Sequences

Arne Skerra, Ph.D., Professor of Biological Chemistry, Technical University Munich

Genetic fusion with conformationally disordered homo-amino-acid polymer sequences provides an alternative way to attach a solvated random chain with large hydrodynamic volume to the protein of biopharmaceutical interest. Such sequences confer high solubility, are resistant against serum proteases, and lead to a large apparent size of the fusion protein. Moreover, they permit efficient production of biochemically active fusion proteins in E. coli. Investigation of the plasma half-life for correspondingly modified antibody fragments and cytokines reveals a pronounced prolongation effect depending on the length and composition of the polymer chain. In particular, genetic fusion with sequences comprising Pro, Ala, and Ser residues can yield pharmacokinetic properties similar to PEGylation, yet without necessitating costly and laborious in vitro modification steps. This PASylation strategy should be useful for the development of therapeutic single-chain and single-domain antibody fragments.

Sponsored by

2:40 

Quartz Crystal Microbalance Biosensors in Immunology Anti-Malarial Antibody Affinity/Avidity Issues
Lars M. Jørgensen Ph.D., University of Copenhagen, Institute for Medical Microbiology and Immunology
Malaria is widespread in tropical and subtropical regions, including parts of the Americas, Asia, and Africa. Each year, there are approximately 515 million cases of malaria, killing between one and three million people; the majority of whom are young children. Malaria is commonly associated with poverty, while it also positively stimulates poverty to become a major hindrance to economic developments. Malaria has currently become one of the most common infectious diseases and an enormous public health problem. Although some vaccines are under development, there is currently none available for malaria. Current preventive drugs must be taken continuously to reduce the risk of infection. Here we use the Attana Quartz Crystal Microbalance technology, as an innovative tool to better understand the mechanism and to improve the development of anti-malarial antibodies.

3:10         Refreshment Break in the Exhibit Hall

4:00         Critical Role of Somatic Mutations in High Potency Human Monoclonal Antibodies to Viruses

James Crowe, M.D., Ingram Professor, Pediatrics and Microbiology

and Immunology, Vanderbilt University Medical Center

Our laboratory has developed novel approaches to development of human monoclonal antibodies that rapidly yield high potency antibodies against viruses. We have isolated potent human monoclonal antibodies from human B cells directed to major human pathogens, including RSV, 1918 and H5 avian influenza, rotavirus, HIV and others. Genetic, biochemical and structural studies reveal that the principal driving force for optimal development of neutralizing human monoclonal antibodies is somatic hypermutation.

4:30         A Novel HPLC-UV-MS Method for Quantitative Analysis of Protein Glycosylation

Anton S. Karnoup, Ph.D., Analytical Sciences, The Dow Chemical Company

5:00         Close of Day

Friday, April 10

Accelerating the Process

7:45         Continental Breakfast in the Exhibit Hall

8:30am   Chairperson’s Remarks

9:05         Selection of Antibody Producing Cells by Toxine Conjugates

Katrin Messerschmidt, Ph.D., Biotechnology, University of Potsdam

The generation of antibodies with designated specificity comprises cost-intensive and time-consuming screening procedures. Here we present a new method by which hybridoma cells can be selected based on the specificity of the produced antibody by the use of antigen-toxin-conjugates thus eliminating the need of a screening procedure. Initial experiments were done with several low molecular weight toxins and fluorescein as model antigen. Several fluorescein-toxin-conjugates were generated, purified by HPLC and characterized regarding their toxicity.

First results show that hybridoma cells that produce fluorescein specific antibodies are able to dispose fluorescein-toxin-conjugates.

9:35         Lead Identification and Optimization in Crude Samples using Label-free Resonant Acoustic Profiling

Helge Schnerr, Ph.D., Product Manager, TTP LabTech, TTP LabTech Ltd.

Although optical-based systems dominate the biosensor market, piezoelectric and acoustic devices represent similar but significantly less expensive alternatives. Acoustic biosensors have been employed in the label-free detection of an incredibly broad range of analytes; from interfacial chemistries and lipid membranes to small molecules and whole cells. Resonant Acoustic Profiling (RAP) technology offers label-free, real-time analysis of biomolecular interactions. The analytical capabilities of a RAP-based biosensor are ideally suited to the development of biotherapeutics, a rapidly expanding area of drug research. In particular, direct measurement in crude and complex samples such as cell culture media or periplasmic extracts eliminates expensive time-consuming purification of often limited material while delivering high content information.

10:05       Coffee Break in the Exhibit Hall

11:05       Cooperation of Dendritic Cells with Naive Lymphocytes on Artificial Extracellular Matrix to Accomplish in vitro Stimulation of Immune Response

Katja Heilmann, Ph.D., Junior Research Group Leader, Biotechnology, University of Potsdam

To serve as a basis for an in vitro immunization a cell culture model was established which combines an artificial extracellular matrix of the paracortical area of lymph nodes with involved immune cell populations of Ova-TCR-transgenic mice and defined antigen. The interaction of dendritic cells, T cells and B cells was investigated by morphological studies, immunfluorescence staining and secretion of interleukins and antibodies. Ovalbumin-specific antibodies could be detected in the culture supernatants after co-cultivation of naive B cells with T cells and dendritic cells. These studies indicate, therefore, that modifications of cell culture substrata with extracellular matrix components support a stronger in vitro interaction of immune cells than plain surfaces and that this effect can be used for the induction of the production of specific antibodies in vitro.

11:35       Pre-Clinical Selection of Non-Immunogenic Lead Antibodies by the Accurate Detection of T Cell Epitopes

Matthew Baker, Ph.D, CSO, Antitope, Ltd.

Helper (CD4+) T cell epitopes are drivers of immunogenicity through induction of class-switched, somatically mutated high affinity antibodies which can bind to and neutralize the injected therapeutic.  Various methods have been developed to detect T cell epitopes in the sequence of therapeutic antibodies.  Several groups have claimed correlation of high affinity MHC class II binding peptides (pMHC) detected using in silico methods with the presence of T cell epitopes, and have proposed the removal of such pMHC. Data will be presented to demonstrate that this approach is flawed as some high affinity pMHC can induce immunological tolerance, whilst certain low affinity pMHC result in effector CD4+ T cell responses that can mediate unwanted immune reactions.

12:05pm Luncheon Presentation I (Opportunity Available)

12:35       Luncheon Presentation II (Opportunity Available)

1:05         Break

CASE STUDES

1:25         Chairperson’s Remarks

1:30         Development and Characterization of Monoclonal Antibodies to Equine Luteinizing Hormone for Diagnostic Applications

Nathalie Forster, M.S., Product Development and Support Manager, Research and Development, Maine Biotechnology Services, Inc.

The success of a hybridoma antibody development project is contingent upon many factors.  Among these factors include the integrity of immunizing and screening reagents, the ability of the fusion screening strategy to discern fusion products of interest, and the capability to characterize the resulting antibodies for use in the end application.  In an effort to generate monoclonal antibodies to equine luteinizing hormone(eLH) suitable for the veterinary reproductive diagnostic market, MBS recently embarked on a hybridoma development project.  Data from this hybridoma project will be presented as well as insight into the development strategies that lead to the successful production of several high affinity, well-characterized monoclonal antibodies.

2:00         High Cytotoxic mAb for the Treatment of FMAI

Margarita Salcedo, Ph.D., Head Scientific Strategy, Medical and Scientific Affairs, LFB S.A.

The LFB has developed a proprietary antibody production process leading to the generation of monoclonal antibodies with high ADCC activity and enhanced affinity for CD16, correlated to its glycosylation pattern. A first generation anti-RhD candidate (R297) has been studied in human healthy volunteers. This monoclonal has been the first monoclonal antibody shown to clear RhD+ red blood as efficiently as polyclonal anti-RhD antibodies in humans. The new generation anti RhD (LFB-593)product will enter into phase  I  clinical trial in Q4 2008.

2:30         Networking Refreshment Break

3:00         Generation of Antibodies Against Toxins

Diana Pauly, Ph.D., Post Doctoral Researcher, Center for Biological Safety, Robert Koch-Institute

The task of raising antibodies against toxins is fraught with one major obstacle: the toxicity of the toxins. In order to ensure survival of the immunized animals, several inactivation strategies have been used to address this problem. The traditional methods are time-consuming and to a variable degree labour-intensive. Therefore, we developed a novel immunization strategy: we covalently immobilized the native toxins onto microbeads in order to reduce their toxicity. We showed that ricin coupled to microbeads is at least 70 times less toxic than native protein in an in vitro cytotoxicity assay and the median lethal dose in mice was 12 times higher for the immobilized toxin than for the native protein. With this new technique we were able to generate highly specific and highly sensitive monoclonal and polyclonal antibodies against native toxins. The antibodies were either used for sandwich immunoassays, multiplex detection systems or functional blockade of the toxin.

3:30         Use of the ClonePix-FL in a Research Monoclonal Facility

Robin Barbour, BS, Associate Director Research, Biology, Elan Pharmaceuticals

The Genetix ClonePix FL™ is an automated fluorescence-based colony selector that allows for high throughput screening of fusions and single step identification of stable single cell clones. We have implemented the use of ClonePix Fl™ to increase our fusion productivity by approximately 50%, while decreasing our time from fusion to stable clone by 50%. The additional benefits of implementing this technology are rescue of previously lost clones, simple selection of high secreting clones and decreased potential of repetitive motion injury to our staff.

4:00         Targeting ED-B Fibronectin: Using BC1-IL12 (AS1409) in Melanoma and Renal Cell Carcinoma

Ben Doran, M.Sc., Research Scientist, Protein Production, Antisoma Research Limited

Clinical responses have been observed to human interleukin-12 in phase I and Phase II trials but development of this molecule was abandoned due to systemic toxicity. It is hoped that targeting this activity by genetically fusing interleukin-12 to a humanised BC1 antibody binding the oncofetal splice variant of fibronectin, extra domain B (ED-B), that reduced side effects and an increased therapeutic index can be observed. Immunohistochemistry shows ED-B to be expressed in renal cell carcinoma and metastatic melanoma which are tumors that have observed clinical efficacy in previous trials involving naked interleukin-12, providing a clear rationale for development.  An efficient manufacturing process with high productivity, unusual for antibody fusion proteins, and high recovery has enabled Antisoma to initiate a phase I dose escalation study in these indications.

4:30         End of Monoclonal Antibodies Conference

For questions or suggestions about the meeting, please contact:
Elizabeth Lamb
Cambridge Healthtech Institute
Phone: 207-493-7874
email: elamb@healthtech.com 

For sponsorship and exhibit information, contact:
Carol Dinerstein
Phone: 781-972-5471 
email: dinerstein@healthtech.com