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）：本研究的目的是开发基于表面增强拉曼散射（SERS）检测的新一代超高通量筛选（uHTS）系统。该项目的具体目标是： 目标 1. 开发多光谱拉曼系统 (MRS) 作为 uHTS 的光学读出装置。目标 2：使用分子哨兵评估 uHTS-MRS，以实现高度并行的配体/靶标结合检测。 目标 3：开发并集成一个自动化系统，用于检测基于 SERS 的纳米探针的光信号。 目标 4：评估 CMOS-uHTS 系统在细胞模型中高通量筛选小分子或 mRNA 探针的情况。 我们的系统将为标签多重分析提供一个新的维度，用于在基于细胞的高通量药物发现筛选中表征大量样品。一种新型的基于 SERS 的分子哨兵检测方案将应用于多重基因表达分析。该系统的独特功能包括： 基于多光谱成像检测的新颖仪器 基于分子识别和 SERS 检测相结合的新颖检测方案 SERS 效应将传统拉曼信号的灵敏度提高了 106-1015 倍 SERS-MS 纳米探针对光漂白不敏感（通常与荧光探针有关）。超高通量“标记多重”检测（即能够同时使用许多拉曼染料标记探针）（极窄的拉曼带允许 SERS 用作多重技术）。 该系统将用于筛选上调或下调各种 mRNA 家族表达的小分子、合成化学品和天然产物库。将鉴定能够调节 mRNA 基因表达水平的活性化合物，然后进行额外的测定。基于 SERS 的 uHTS 仪器将提供一种工具，该工具有可能比当前可用的系统更快、更高效，并且能够提供高灵敏度和更高水平的特异性，以及筛选合成化学和天然产物库的多重功能，例如将在 NIH 赞助的分子图书馆筛选中心注册和安置。