High Throughput Modular Microfluidic Systems for Drug Discovery/Development

用于药物发现/开发的高通量模块化微流体系统

• 批准号: 7900284
• 负责人: Steven Allan Soper
• 金额: $ 11.84万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7900284
• 关键词: Aging; Area; Biochemical; Biological; Biological Assay; Bioreactors; Cell Aging; Cells; Characteristics; Communities; Consensus Sequence; Consumption; Data; Data Quality; Detection; Development; Disease; Dyes; Elements; Energy Transfer; Enzymes; Equipment; Event; Fluorescence; Fluorescence Resonance Energy Transfer; Fluorescent Probes; Fluorochrome; Generations; Genetic; Genome; Human Genome; Image; Immobilization; Indium; Intervention; Kinetics; L1 Elements; Label; Libraries; Liquid substance; Malignant Neoplasms; Mediating; Metals; Methods; Metric; Microfluidics; Molds; Molecular; Monitor; ORF2 protein; Oligonucleotides; Optics; Performance; Phase; Play; Polymers; Population; Process; Reaction; Reagent; Recycling; Reporting; Research; Research Personnel; Retrotransposition; Robotics; Role; Sampling; Scheme; Screening procedure; Solid; Source; Spectrum Analysis; Speed; Structure; Surface; System; Technology; Testing; Work; base; cancer therapy; charge coupled device camera; combinatorial; cost; detector; drug candidate; drug discovery; endonuclease; enzyme activity; enzyme substrate; feeding; flexibility; high throughput screening; human disease; improved; inhibitor/antagonist; innovation; models and simulation; next generation; novel; parallel processing; phthalocyanine; programs; quantum; response; submicron; therapeutic target

DESCRIPTION (provided by applicant): BioMEMS (Biological-MicroElectroMechanical Systems) are seen as the next generation platform for performing many different biologically-based assays in areas such as drug discovery, due to their potential for providing improved performance and low assay cost. However, in reports to date of BioMEMS used in high throughput screening (HTS), the potential improvements in throughput and costs have yet to be realized. This research focuses on developing new highly integrated polymer BioMEMS platforms to achieve advantages in the screening of libraries of drug candidates using parallelized infrared fluorescence assays. As a demonstration of the efficacy of the technology, inhibitors of L1 endonuclease (L1-EN) will be screened. L1-EN induces double-strand breaks via a TTTT'AA consensus sequence and can contribute to genetic damage responsible for cellular aging, cancer formation or progression. Therefore, discovery of inhibitors of L1-EN, of which there is currently none, could play an important role in minimizing genetic instability during cancer therapies or certain aspects of aging. The platforms to be developed have several enabling technologies that will allow the dissemination of HTS capabilities into the broader drug-discovery community by significantly reducing equipment demands and minimizing consumable costs. The system will consist of a large number of fluidic processors (96-192) configured on a 6" polymer wafer produced using micro- replication. Each fluidic processor will consist of (1) interconnected chip(s) to feed compounds directly from titer plates to the main processor wafer; (2) micromixer that speeds reagent mixing; (3) high surface area bioreactor in which the target is immobilized; and (4) grouped fluidic flow-cells for identifying potential leads using multi-channel fluorescence detection. A substrate of L1-EN will be dual-labeled with new, near-IR fluorescent phthalocyanine dyes, which provide high sensitivity and low background. Parallel fluorescence readout from the fluidic flow cells will be carried out by imaging a group of tightly spaced channels onto a CCD camera. Simulation and modeling will guide the selection of an appropriate readout format and optical configuration. In its finished format, the system will be able to screen -190,000 drug candidates in 24 hr. Fluid handling will be accomplished via microfluidics with minimal robotic intervention, as only a single sample transfer step is required to load the system with drug candidates.

描述（由申请人提供）：BioMEMS（生物微机电系统）被视为在药物发现等领域执行许多不同的基于生物学的测定的下一代平台，因为它们具有提供改进的性能和较低的测定成本的潜力。然而，在迄今为止用于高通量筛选 (HTS) 的 BioMEMS 的报告中，通量和成本的潜在改进尚未实现。本研究重点开发新型高度集成的聚合物 BioMEMS 平台，以利用并行红外荧光测定在筛选候选药物库方面取得优势。为了证明该技术的功效，将筛选 L1 核酸内切酶 (L1-EN) 抑制剂。 L1-EN 通过 TTTT'AA 共有序列诱导双链断裂，并可能导致导致细胞衰老、癌症形成或进展的遗传损伤。因此，L1-EN 抑制剂（目前还没有）的发现可能在最大限度地减少癌症治疗或衰老某些方面期间的遗传不稳定性方面发挥重要作用。待开发的平台拥有多项支持技术，可通过显着减少设备需求并最大限度地降低消耗品成本，将 HTS 能力传播到更广泛的药物发现领域。该系统将由大量流体处理器（96-192）组成，配置在使用微复制生产的6英寸聚合物晶片上。每个流体处理器将由（1）互连芯片组成，用于将化合物直接从滴定板供给到主处理器晶片；（2）加速试剂混合的微混合器；（3）其中固定靶标的高表面积生物反应器；以及（5）生物反应器。 (4) 分组流体流动池，用于使用多通道荧光检测来识别潜在的引线。 L1-EN 的底物将用新型近红外荧光酞菁染料进行双重标记，从而提供高灵敏度和低背景。流体流动池的并行荧光读数将通过将一组紧密间隔的通道成像到 CCD 相机上来进行。模拟和 建模将指导选择适当的读出格式和光学配置。在最终的格式中，该系统将能够在 24 小时内筛选 -190,000 种候选药物。流体处理将通过微流体技术以最少的机器人干预来完成，因为只需要单个样品转移步骤即可将候选药物加载到系统中。