Protein Production for StructureBased Drug Design基于结构的药物的设计的蛋白质的生产.pptVIP

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Protein Production for StructureBased Drug Design基于结构的药物的设计的蛋白质的生产.ppt

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Protein Production for StructureBased Drug Design基于结构的药物的设计的蛋白质的生产

Vertex: Building a Major Drug Company Focus:Small molecule drugs for major diseases Established: 1989; Public: 1991 Common stock: NASDAQ: VRTX 700 employees 3 sites Cambridge, US (headquarters) Oxford, UK San Diego, CA Traditional vs. Vertex Approach to Drug Discovery Therapeutic Area Perspective Single target approach Target Family Perspective Chemogenomics multi-target approach Expanding Parallel Drug Discovery into Gene Families Content Structural Genomics vs Structure Based Drug Design Much in common Expression strategy for higher output Parallel expression of E.coli insect cells Quantitative analysis Illustrated using examples from Vertex Kinase program Application to other protein families Integration into a broader process with other disciplines Structure Based Drug Design Drug Discovery ( Development) Human proteins Complex post-translationally modified proteins Heterogeneous proteins ‘Difficult’ proteins Highest value structures contain inhibitor Premium given to high-output (cf high-throughput) Failure not an option Protein Expression Bottleneck Expression Process Triage Expression Prior to Production: Expression systems Growth conditions Cell lines Constructs Mutants pBEV: Dual-System Expression Vector Expression Systems Highly Engineered Process Using Standard Equipment HT-Expression Automated Purification Full-Length Kinases in E.coli Insect cells Model Behavior in E.coli Effect of Protein Size on Expression Solubility in E.coli Quantitative Analysis of Expression Strategies:Decision Tree Analysis of Kinase Expression Novel Kinase Structures in PDB Crystal Structure of MAP Kappa 2 Kinase Crystal Structure of Aurora-2 Kinase Crystal Structure of FLT-3 Kinase HT-Expression Allows the Exploration of Diversity:Mutations HT-Expression Allows the Exploration of Diversity:Utilizing Different Cell Lines Insect Cell Expressed Structures in PDB E.coli Expressed Structures in PDB From the Vertex Portfolio Expressing Difficult Proteins in I