Microarraying and Screening Chemical Libraries

微阵列和筛选化学文库

• 批准号: 7546586
• 负责人: HAICHING MA
• 金额: $ 43.54万
• 依托单位: REACTION BIOLOGY CORPORATION
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-13 至 2010-05-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7546586
• 关键词: Aerosols; Antigens; Archives; Automation; Back; Biochemical; Biological Assay; Biology; Biotin; Buffers; Caspase; Chemicals; Data; Deposition; Detection; Development; Dimethyl Sulfoxide; Engineering; Environment; Enzymatic Biochemistry; Enzyme-Linked Immunosorbent Assay; Failure; Fluorescence; Gel; Glass; Glycerol; Goals; Head; Humidity; Image; Individual; Inhibitory Concentration 50; International; Lasers; Libraries; Liquid substance; Maps; Matrix Metalloproteinases; Mechanics; Microfabrication; Miniaturization; Molecular Bank; Peptide Hydrolases; Phase; Phosphotransferases; Plastics; Printing; Process; Production; Productivity; Protein Binding; Protein Inhibition; Protein Tyrosine Kinase; Protein-Serine-Threonine Kinases; Proteins; Protocols documentation; Reaction; Reagent; Recombinant Proteins; Research Personnel; Resolution; Running; Scanning; Screening procedure; Serine Protease; Shipping; Ships; Site; Speed; Spottings; Streptavidin; Surface; System; Technology; Testing; Training; Tube; Ultrasonics; United States National Institutes of Health; Validation; assay development; base; cofactor; design; design and construction; detector; drug discovery; enzyme substrate; experience; high throughput screening; inorganic phosphate; instrument; instrumentation; large print; meter; micro-total analysis system; nanoDroplet; nanolitre; nanolitre scale; programs; protocol development; repository; small molecule libraries; software development; tool

Reaction Biology Corporation (Malvern PA) has developed homogeneous-phase reaction microarrays to serve the NIH Roadmap initiative for nanoliter-scale biochemical high throughput screening (HTS) in drug discovery, large scale IC50 determinations, and selectivity profiling. These nanoliter reactions are 1000 to 10,000-fold smaller than well plate formats currently in use. Preliminary data is presented for microarray HTS against serine proteases, caspases, MMPs, tyrosine kinases, and serine/threonine kinases. Aim 1: A chemical microarrayer will be designed and built through a Consortium with TeleChem (Sunnyvale CA). This microplate arrayer will deploy a 384-pin manifold to print NIH library compounds to array-plates containing up to 38,400 compounds per glass array-plate (SBS standard). The design goal for this instrumentation is to achieve the printing of 10 sets of the entire NIH repository in a 24-hr print run to be used for 10 individual HTS campaigns (5 million reactions). Aim 2: An array-plate activator will be designed and validated for the rapid delivery of sub-nanoliter quantities of enzymes, substrates, cofactors, and detectors to the compound array-plates. The design goal for this instrumentation will be to initiate biochemical screens against the entire NIH compound repository (20 array-plates containing 500,000 compounds) in 30 minutes. Aim 3: Construction and delivery of an array-plate activator beta-unit to the NIH for testing and validation. This instrumentation seeks to exploit the tremendous liquid handling power of microarrayers, the miniaturization of microarrays for compound conservation and easy HTS assay development, and the resolution/sensitivity of multicolor laser scanners. Other applications are enabled by this instrumentation: (1) the rapid printing of large chemical libraries on plates pre-coated with an immobilized protein and binding partner would facilitate protein-protein inhibition HTS; (2) the ability to rapidly print 100,000 or more cellular lysates produced in well plate HTS facilitates highly multiplexed detection of numerous intracellular antigens by reverse phase arrays analyzed by sandwich ELISA. Overall, this instrumentation allows a large number of wild type or recombinant protein targets, each obtained in miniscule quantity, to be screened against 100,000 to 500,000 compounds of the NIH repository.

反应生物学公司（马尔文PA）已经开发了反相反应微阵列， 服务于NIH的纳米级生化高通量筛选（HTS）药物路线图倡议 发现、大规模IC 50测定和选择性分析。这些纳升反应是1000到 比目前使用的孔板格式小10，000倍。初步数据为微阵列 HTS针对丝氨酸蛋白酶、半胱天冬酶、MMP、酪氨酸激酶和丝氨酸/苏氨酸激酶。目标1：A 化学微阵列将通过TeleChem（桑尼维尔CA）的联合体设计和制造。这 微板阵列将部署一个384针歧管打印NIH库化合物阵列板含有 至38，400种化合物/玻璃阵列板（SBS标准）。该仪器的设计目标是 在24小时打印运行中实现整个NIH存储库的10套打印，用于10个个体 HTS活动（500万个反应）。目标2：将设计并验证阵列板激活器， 快速递送亚纳升量的酶、底物、辅因子和检测剂至化合物 阵列板。这种仪器的设计目标是启动针对整个生物化学的筛选。 NIH化合物储存库（20个阵列板，含有500，000种化合物）。目标三： 构建阵列板激活剂β-单元并将其交付给NIH进行测试和验证。 该仪器试图利用微阵列的巨大液体处理能力， 微阵列的小型化，用于化合物保存和易于HTS测定开发，以及 激光扫描仪的分辨率/灵敏度。该仪器还支持其他应用：（1） 在预先涂有固定化蛋白质的板上快速打印大的化学文库， 伴侣将促进蛋白质-蛋白质抑制HTS;（2）快速打印100，000或更多细胞的能力 在孔板HTS中产生的裂解物有助于多种细胞内抗原的高度多重检测 通过夹心ELISA的反相阵列分析。总的来说，这种仪器允许大量的 的野生型或重组蛋白靶，每一个都以极小的量获得， 100，000到500，000种化合物。