- Pfizer
- GlaxoSAE Media GroupthKline
- Bayer-Schering Pharma
- Roche Products Ltd
- Merck Research Labs
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Conference agenda
High-throughput production of eukaryotic proteins for X-ray crystallography and NMR spectroscopy structure determination Ronnie Frederick, Protein Production Team Leader , Center for Eukaryotic Structural Genomics, University of Wisconsin Expression vector design and E.coli Auto-induction system for protein productionAutomated analysis of Protein and Domain production (Caliper LC90) Meso- and Small-scale Semi-automated Protein purification (Maxwell 16 system)Time and cost considerationsFuture developments in Eukaryotic Protein productionConclusion Developing the bacterial cell factory: Quiescent E. coli for recombinant protein expression David Summers, Head of Department, Cambridge University Stability functions of bacterial multicopy plasmidsPlasmids that control the bacterial cell cycleExploiting cell cycle control to achieve bacterial quiescenceProtein expression in quiescent cellsReinventing the system; second generation quiescent cells Optimised pipeline purification of over-expressed eukaryotic proteins Karl Nichols, Research Specialist, University of Wisconsin Overview of the Centre For Eukaryotic Structural Genomics’ Purification ProcessQuality Control of the Purification Process Successes of the Purification ProcessDiscussion of Purification FailuresSalvage Pathways: Attempting to Rescue the Failures Development and application of high throughput methods for structural & functional studies of membrane proteins Vincent Postis, Research Fellow, University Of Leeds Vector design and E. coli Auto-induction system Automated analysis of membrane protein and soluble domain productionHigh-throughput membrane protein solubilisationHigh-throughput protein stability assessment Furthering membrane protein structural and functional studies using cell-free expression technologies Michael Goren, Research Assistant, University Of Wisconsin-Madison Limitations of in vivo expression: Cell-Free vs Cell-Based Cell-free expression for NMR structural analysis of membrane ProteinsDirect cell-free incorporation into liposomes and activity of Membrane ProteinsIn vitro reconstitution of membrane protein systems HT-biochemical annotation of genetic information using the wheat germ cell-free system Yaeta Endo, Executive Director, Ehime University The wheat gem cell-free protein production system Applications to genome-wide functional and structural analyses Application to difficult protein expression (multi-subunit proteins, membrane proteins) New robotic system for the massive protein production High-throughput crystallomics: Translating technology from the protein structural initiative to the biomedical research community Craig Bingman, Section Leader, Crystallography and Bioinformatics, University Of Wisconsin-Madison Architectural criteria for appropriate crystallomics automationCESG’s extensible platformReview of crystallomics results from the protein structural initiativeAdditive screening results Discussion & Questions – Review of the Session Ronnie Frederick, Protein Production Team Leader , Center for Eukaryotic Structural Genomics, University of Wisconsin Close of Executive Briefing Chairman's Opening Remarks Bernardo Perez-Ramirez, Scientific Director, BioFormulations Development, Genzyme Corporation GENE SYNTHESIS IN PROTEIN ENGINEERING Mark McAlister, Associate Director, Protein Engineering, Global Structural Chemistry, AstraZeneca The role of protein engineering Conventional molecular biology vs. gene synthesisCase studiesFuture directions USE OF LABCHIP 90 IN PRODUCT DEVELOPMENT OF A RECOMBINANT FUSION PROTEIN Joseph Siemiatkoski, Senior Scientist, Biogen Idec Method development timelines can be compressed using automationSpeed of analysis and high throughput are crucial for development effortsWide linear range simplifies operability and data analysisFor quantitation, selectivity of separation has benefits over traditional approachesData quality is improved by enabling replicate analysis – statistically justified purity data STRUCTURE-FUNCTION STUDIES OF INTERLEUKIN-21 Kent Bondensgaard, Research Scientist, Novo Nordisk 3D structure of IL-21Conformational heterogeneityReceptor interactionsDesign of superpotent IL-21 analoguesMechanisms for increased potency by cytokine analogues EXCELLENCE IN DNA ENGINEERING AND PROCESSING Nina Albrecht, Manager, Geneart AG Expression optimisation – background, rationales and GENEART philosophyThe art of optimising genes: a multi-parameter challengeCase studies of different expression systems pre and post optimisation DNA engineering and processing workflowImproved proteins by directed evolutionIndresh Srivastava, Head of Protein Biology, Novartis Vaccines and Diagnostics Select an appropriate expression systemsStrategies for selecting an appropriate structure for clinical evaluationProduction of selected vaccinePurification of vaccine immunogenCharacterization of the vaccine using state of the art techniques for determining the conformation; glycosylation, epitope exposure, binding affinity, and stabilityClinical evaluation of vaccines COMBINATORIAL DOMAIN HUNTING IN MOSAIC MODE Renos Savva, Senior Research Fellow, Birkbeck College, University of London The provision of soluble, folded and stable recombinant proteinCombinatorial Domain Hunting (CDH) technologyAssessing time and resource effective approaches to expression construct designIs it necessary to access bioinformatics analysis in construct making?Provision of previously recombinantly intractable/ insoluble proteinsThe isolation of protein-protein complexes in soluble, stable recombinant arrangementsOptimising recombinant expression for structure and drug design CASE STUDY: MULTI-PARALLEL PROTEIN PRODUCTION Ian Hunt, Group Leader, Novartis Utility of multi-parallel E. coli baculovirus mediated protein expression strategiesCurrent bottlenecks within the processPotential new and enabling technologiesMark Abbott, Principal Scientist, Respiratory and Inflammation, AstraZeneca Multi-domain proteinsMethods to handle and analyse different post translational modificationsMammalian expression with many different ‘flavours’Easy access to mammalian, insect and E.coli expression systems essential for successDifferent expression systems for different usesCase studies of different protein families PROTEIN SEPARATION AND BIOMARKER DISCOVERY USING LABEL FREE INTRINSIC IMAGING Judit Nagy, Proteomics Research Scientist, Imperial College London Optimising the processMethods usedValidation of marker candidatesProteomic technologies to find novel markers CELL-FREE EXPRESSION OF INTEGRAL MEMBRANE PROTEINS USING DISCOIDAL MEMBRANES W. Antoni Kudlicki, Research Fellow, R&D, Invitrogen Discoidal membranes (NLPs)NLP-based expression and solubilisation of membrane proteinsCell-free production of medically relevant NLP-MP complexes using ExpresswayDifferent strategies for complex formationFunctional analysis and ligand displacement assay for selected membrane protein targetsCharacterisation of MP-NLP complexes Chairman’s Closing Remarks and Close of Day One Bernardo Perez-Ramirez, Scientific Director, BioFormulations Development, Genzyme Corporation Drinks Reception sponsored by Caliper LifeSciences Chairman's Opening Remarks Ian Hunt, Group Leader, Novartis ASSESSING THE SOLUTION BEHAVIOR OF THERAPEUTIC PROTEINS Bernardo Perez-Ramirez, Scientific Director, BioFormulations Development, Genzyme Corporation Biophysical approaches to pre-formulationMaximizing data out put with minimal amounts of protein Global analysis and thermodynamic assessment Choosing proper stress conditions PROTEIN EXPRESSION IN INSECT CELLS Linda King, Director, Oxford Expression Technologies Ltd New vectors for multi-parallel protein productionIncreasing yields and quality of difficult to express proteinsRapid titration of baculovirus expression vectorsMicro scale and large scale protein production A NOVARTIS CASE STUDY: KINASE AND PROTEASE PREPARATION Paul Ramage, Senior Research Investigator 1, CPC/Protease Platform, Novartis Pharma AG Overview/organisationProblems facedSolutionsSpecific examplesFuture directions ENGINEERING ENHANCED THERAPEUTIC PROTEINS Protein engineering technologiesExamplesOptimisation of therapeutic proteins and antibodiesBasis of protein immunogenicityEarly assessment of protein immunogenicityImmunogenicity engineering Ingrid Marchal-Gerez, Business Development Manager, Proteus Laurent Fourage, Project Leader, Biotechnology, Proteus NOVEL TOOLS FOR HIGHER THROUGHPUT AND HIGHER PROTEIN YIELD IN MAMMALIAN AND INSECT CELLS Balwant Patel, Director, Market Development, Cloning and Protein Expression Buisness Unit, Invitrogen Multi-site Gateway® Pro, simultaneous cloning of up to four gene fragments into any Gateway® expression vector.pFastBac™ TOPO and BaculoDirect™ systems for rapid high throughput insect expressionFreeStyle™ MAX System, a gram of protein from 293 or CHO cells in just one weekHigh efficiency gene delivery into almost impossible to transfect cell types with ViraPower HiPerformTM Lentivirus Expression System LARGE SCALE PROTEIN PRODUCTION Florian Wurm, Professor of Biotechnology, Ecole Polytechnique Federale De Lausanne Multi-gram per litres - how to achieve thisDNA to process in the shortest timeGenetics and the biology of large scale manufacturing Transient gene expression – the innovative new approachDisposables at large scale? HIGH THROUGH PUT PROTEIN PURIFICATION Gisela Schnapp, Principal Scientist, Boehringer Ingelheim Expression screening with small amount of proteinHigh through put protein purification methods for crystallisation trialsprotein characterisation Purity versus yieldTime and cost implicationsCrystallisation methods PROTEOMICS-BASED VALIDATION OF GENOMIC DATA Ignacio Casal, Biotechnology Programme Director and Head of the Protein Technology Unit, CNIO Parallel expression of multiple proteinsEvaluation of different approaches for antibody productionPhage display and PrEST technologyValidation strategies for tumoral biomarkers STRUCTURE DETERMINATION OF PROTEIN-LIGAND COMPLEXES Rick Davies, Team Leader, AstraZeneca Co-expression with ligandsSurface mutagenesisConstruct designHybridisationUse of orthologues CHECKPOINT RESEARCH IN MITOSIS Peter Hemmerich, Research Associate, FLI Leibniz Institute for Age Research Effective cloningIn vivo fluorescence techniques (FRAP, FCS, FCCS, FRET, FLIM)Structural architecture of the kinetochoreDynamics of kinetochore proteinsExemplary approach – its application to other cellular machineries Chairman’s Closing Remarks and Close of Conference
Workshops
Copthorne Tara Hotel 4 June 2008 London, United Kingdom
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