Ultra-High-Throughput Screening (uHTS) Based on Surface*

基于表面的超高通量筛选 (uHTS)*

• 批准号: 7318325
• 负责人: Tuan Vo-Dinh
• 金额: $ 46.84万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-02 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7318325
• 关键词: Binding; Biological; Biological Assay; Biological Factors; Cell model; Cells; Chemicals; Detection; Dimensions; Dyes; Family; Fluorescent Probes; Gene Expression; Generations; Housing; Image; Label; Libraries; Ligands; Messenger RNA; Molecular; Molecular Bank; Numbers; Optics; Pathway Analysis; Photobleaching; Range; Research; Sampling; Scheme; Screening procedure; Sentinel; Signal Transduction; Specificity; Surface; System; Techniques; Testing; Tissues; base; design; drug discovery; high throughput screening; instrumentation; molecular recognition; multiplex detection; nanoprobe; novel; small molecule; tool

DESCRIPTION (provided by applicant): The objective of this research is to develop a new generation of ultra-high-throughput screening (uHTS) systems based on surface-enhanced Raman scattering (SERS) detection. The specific aims of the project are: Aim 1. Develop a multi-spectral Raman system (MRS) as the optical readout for uHTS. Aim 2: Evaluate the uHTS-MRS using molecular sentinels for highly parallel ligand/target binding detection. Aim 3: Develop and integrate an automated system for detection of optical signals from SERS-based nanoprobes. Aim 4: Evaluate the CMOS-uHTS system for high throughput screening of small molecules or mRNA probes in a cellular model. Our system will offer a new dimension in label multiplexing for characterizing large numbers of samples in cell-based high-throughput screening for drug discovery. A novel SERS-based molecular-sentinel detection scheme will be applied for multiplex gene expression analysis. The unique features of the proposed system include: Novel instrumentation based on multi-spectral imaging detection Novel detection scheme based on combination of molecular recognition and SERS detection The SERS effect enhances the sensitivity of conventional Raman signal over 106-1015 fold SERS-MS nanoprobes are insensitive to photo-bleaching (often associated with fluorescent probes). Ultra high-throughput "label multiplex" detection (i.e. capability for many Raman dye-labeled probes to be used simultaneously) (the extremely narrow Raman bands allow SERS to be used as a multiplexing technique). The system will be tested for screening of small molecules, synthetic chemical and natural product libraries that up- or down-regulate expression of various families of mRNA. Active compounds that are capable of modulating mRNA gene expression levels will be identified, followed by additional assays. The SERS-based uHTS instrumentation will provide a tool that has the potential to be faster and more efficient than currently available systems, and is capable to provide great sensitivity with higher levels of specificity, and multiplexing capabilities to screen synthetic chemical and natural product libraries such as the ones that will be registered and housed in the NIH-sponsored molecular libraries screening centers.

描述（由申请人提供）：这项研究的目的是基于表面增强的拉曼散射（SERS）检测开发新一代的超高通量筛选（UHTS）系统。该项目的具体目的是：目标1。开发多光谱拉曼系统（MRS）作为UHT的光学读数。 AIM 2：使用分子哨兵评估UHTS-MR，以高度平行的配体/靶结合检测。 目标3：开发和集成一个自动化系统，以检测基于SERS的纳米探针的光学信号。 AIM 4：评估CMOS-UHTS系统在细胞模型中对小分子或mRNA探针的高吞吐量筛选。 我们的系统将在标签多路复用中提供一个新的维度，以表征基于细胞的高通量筛选以发现药物发现的大量样品。一种新型的基于SER的分子 - 塞替奈尔检测方案将用于多重基因表达分析。该系统的独特特征包括：基于多光谱成像检测的新型仪器，基于分子识别和SERS检测的组合，基于多光谱成像检测方案，SERS效应增强了传统拉曼信号在106-1015倍SERS-MS-MS纳米螺旋体上的敏感性，对摄影质量不太敏感（通常与荧光探针相关联）。超高通量“标签多路复用”检测（即，许多拉曼染料标记的探针可以同时使用）（极狭窄的拉曼带允许SERS用作多重技术）。 该系统将进行测试，以筛选小分子，合成化学和天然产物文库，这些库是各种mRNA家族的表达或下调表达的。能够调节mRNA基因表达水平的活性化合物，然后进行其他测定。基于SERS的UHTS仪器将提供一种工具，具有比当前可用系统更快，更有效的工具，并且能够提供具有更高水平的特异性的敏感性，以及筛选合成化学和天然产品库的多重功能，例如将在NIH NIH的分子图书馆筛选中注册和住宿的敏感性。