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1:30 Chairperson’s Opening Remarks
Mitchell Ho, Ph.D., Head, Antibody Therapy Unit, Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health (NIH)
1:40 OPENING KEYNOTE PRESENTATION
Emerging Mass Spectrometry-Based Tools to Characterize Higher Order Structure and Dynamics of Biopharmaceuticals
Igor A. Kaltashov, Ph.D., Associate Professor, Department of Chemistry, University of Massachusetts, Amherst
Among several MS-based techniques, targeting protein higher order structure and dynamics, hydrogen/deuterium exchange (HDX) has demonstrated the greatest promise vis-à-vis conformational analysis of biopharmaceutical products. Several examples of biopharmaceutical products (interferon beta 1a, velaglucerase, etc.) will be used to illustrate the utility of HDX MS and related techniques as a means of characterizing protein drugs in terms of their conformational integrity, stability and functional competence with high predictive value.
2:10 Quantitative Method for Measurement of Antibody Internalization using Fluorescent Imaging
Inna Vainshtein, Ph.D., Scientist II, Global PK-PD & Bioanalysis, MedImmune
Quantitative assessment of internalization is important in development of antibody therapeutics. Despite a number of publications describing antibody-mediated receptor internalization, quantitative assessment of this process has not been extensively presented. Target-mediated internalization may increase antibody clearance and result in non-linear pharmacokinetic (PK) profiles. For immunotoxins, internalization could effect efficiency of toxin delivery into the target cells. We have developed a quantitative image-based method for measurement of antibody internalization. Examples will be presented to demonstrate the application of this methodology to development of therapeutic antibodies.
2:40 Antibody Screening using Multiplexed SPR
John Corbin, Ph.D., XOMA (US) LLC
Understanding how an antibody exerts a therapeutic effect in vivo often depends on knowing the mechanism by which the antibody impacts the targeted signaling pathways at a molecular level. This is especially relevant for XOMA 052, an anti-IL-1β antibody that regulates the activity of the cytokine by a novel mechanism of differentially modulating the kinetic parameters of IL-1β binding to its cognate receptors. Biophysical studies using techniques such as surface plasmon resonance (SPR) are a powerful approach for characterizing therapeutic antibody mechanism of action as well as a facile technique for mechanistic screening of antibodies. Analysis of multiple molecular interactions in parallel using multiplexed SPR offers several advantages over conventional SPR including increasing throughput and the ability to conduct side-by-side comparisons of binding kinetics under different conditions. This presentation will highlight the use of surface plasmon resonance to elucidate antibody mechanism of action
2:55 High Throughput, Small Scale, and Optimization of Protein Separations Using Automated PhyNexus Platform Technology
Lee Hoang, Ph.D., Manager, Research & Development, PhyNexus Inc.
Characterization of therapeutic candidates requires that proteins are well purified post expression. We have developed a platform that completely automates purification, enrichment and desalting of functional proteins eliminating bottlenecks associated with traditional protein purification techniques and expediting multiple stages of the discovery process. Examples of how the platform is utilized in biomarker analysis, process and assay development, and immunogenicity will be presented. Protein separations in small-scale extraction columns with optimized conditions enabling functional and analytical tests will be discussed as well.
3:10 Refreshment Break, Poster and Exhibit Viewing
4:00 Combining Label Free Assay Platforms to Support Therapeutic Antibody Development from Identification of Candidate Antibodies through Pre-Clinical Development
Robin Barbour, Director, Antibody Technology, Elan Pharmaceuticals
Label free technologies can impact antibody development from the earliest phase through entry into the clinic and beyond. In this presentation, we compare and contrast three optically-based label free technologies, the Biacore T100, the Forte-Bio Octet, and SRU bind in their ability to screen antibodies from tissue culture supernatants and then to characterize the resultant positives for affinity, kinetics, epitope binding and domain binding. During the presentation, the advantages and disadvantages of each technology will be highlighted.
4:30 Proteomic Profiling of Novel Protein Targets by Selective Epitope Inhibition and SILAC/MS Analysis
Christian Freund, Ph.D., Principal Investigator/Group leader, Structural Biology, FMP/Free University of Berlin
We have a developed a rapid and robust method for addressing specificity of protein-protein interactions by a combined inhibitor/SILAC/MS approach. As exemplified by intracellular adaptor domains involved in disease processes, we show that deconvolution of epitopes is possible. This allows one to define the contribution of individual interaction sites for the assembly of molecular machines (e.g., the spliceosome) or signaling pathways.
5:00 End of Day
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