Barcoded Hydrogel Microparticles and Scanner for Multiplexed Biomolecule Assays

用于多重生物分子检测的条形码水凝胶微粒和扫描仪

• 批准号: 7659888
• 负责人: Patrick S Doyle
• 金额: $ 22.56万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7659888
• 关键词: Affect; Area; Base Pairing; Binding; Biological; Biological Assay; Blood typing procedure; Cells; Chemicals; Clinical; Code; Complex; Computer software; Cost Savings; DNA; Detection; Development; Devices; Dextrans; Diagnostic; Discrimination; Drug Formulations; Fluorescence; Free Radicals; Gene Expression Profiling; Genomics; Genotype; Goals; Healthcare; Height; Hydrogels; Image; In Vitro; Kinetics; Length; Measures; Medical; Medicine; Metric; Microfluidic Microchips; Microfluidics; Microscopy; Modeling; Molecular; Molecular Weight; Morphology; Nucleic Acids; Performance; Polymers; Process; Property; Proteins; Public Health; Reading; Reproducibility; Research; Route; Safety; Sampling; Scanning; Scheme; Screening procedure; Sensitivity and Specificity; Signal Transduction; Solutions; Specificity; Structure; System; Techniques; Technology; Testing; Time; Transfusion; Ursidae Family; Variant; Work; Writing; base; combinatorial chemistry; density; design; dextran; disease diagnosis; drug candidate; drug development; drug discovery; flexibility; genetic analysis; high throughput screening; light intensity; new technology; next generation; particle; photomultiplier; physical property; prenatal health; public health relevance; success; tool

DESCRIPTION (provided by applicant): Multiplexed screening is a tool that finds broad use in applications such as drug discovery, genotyping, medical diagnostics, and blood typing for transfusion safety, and will be of utmost importance in the up-and-coming field of "personalized" medicine. The two commercially available screening technologies offer either a high "density" of analytes measured (i.e. planar microarrays) or high sample throughput (ie. bead-based systems), but not both. This application proposes the fundamental development of a new screening technology, based on multi-functional encoded particles, which could provide the density of microarrays and throughput of bead-based systems. Preliminary results show that particles composed of a spongy hydrogel material, with a punch-code barcode written on one half and a stripe for target capture on the other, can be used to simultaneously quantify targets in a single biological sample, with coding capabilities of over one million. In building upon a proof-of-concept demonstration, it is hypothesized that (1) increasing the size of pores in the hydrogel structure will allow targets to bind throughout the particle, increasing the sensitivity of each assay while decreasing required incubation times; (2) that hybridization conditions can be tuned to achieve performance competitive with existing technologies; and that (3) a flow-through system based on microfluidics and photomultiplier technologies can be used to rapidly scan particles (i.e. read codes and quantify targets). The specific aims of the project are: (1) Enhance particle synthesis by exploring chemical variations and processing conditions to generate particles that are sufficiently porous and mechanically robust. Particles will be examined via microscopy and probed with FITC-conjugated dextrans. (2) Optimize the physical and chemical conditions of DNA hybridization assays to maximize sensitivity, specificity, and reproducibility. (3) Develop a microfluidic flow-based system for rapid scanning that integrates a flow-focusing microfluidic device, photomultiplier-aided fluorescence detection, and software to decode the acquired signal. The end goal of this project is to have a system capable of quantifying 2,500 nucleic acid targets per sample, detecting targets with single base-pair discrimination at a better sensitivity than commercially available systems, and scanning 5,000 particles at a rate of 500 particles per minute. The relevance of this project to public health is the development of a transformative technology for genomic medicine, ranging from disease diagnosis to drug discovery. PUBLIC HEALTH RELEVANCE: This project will develop a new technology that can be used to simultaneously detect thousands of biomolecules in a solution. This new technology will find potential use in disease diagnosis/treatment, blood typing for increasing the safety of transfusions and drug development.

描述（由申请人提供）：多重筛选是一种广泛用于药物发现、基因分型、医学诊断和输血安全血型分型等应用的工具，在新兴的“个性化”医学领域将具有极其重要的意义。这两种商业上可用的筛选技术提供了高“密度”的分析物测量（即平面微阵列）或高样品通量（即，基于珠粒的系统），但不是两者。本申请提出了一种新的筛选技术的基本发展，基于多功能编码颗粒，它可以提供微阵列的密度和基于珠的系统的吞吐量。初步结果表明，由海绵状水凝胶材料组成的颗粒，其中一半写有穿孔码条形码，另一半写有用于目标捕获的条纹，可用于同时量化单个生物样品中的目标，编码能力超过一百万。在建立概念验证证明的基础上，假设（1）增加水凝胶结构中的孔的尺寸将允许靶结合整个颗粒，增加每个测定的灵敏度，同时减少所需的孵育时间;（2）可以调节杂交条件以实现与现有技术竞争的性能;以及（3）基于微流体和光电倍增管技术的流通系统可用于快速扫描颗粒（即，读取代码和量化目标）。该项目的具体目标是：（1）通过探索化学变化和加工条件来增强颗粒合成，以生成足够多孔和机械坚固的颗粒。将通过显微镜检查微粒，并用FITC结合的葡聚糖进行探测。(2)优化DNA杂交分析的物理和化学条件，以最大限度地提高灵敏度、特异性和重现性。(3)开发一种基于微流控流动的快速扫描系统，该系统集成了流动聚焦微流控装置、光电倍增管辅助荧光检测和软件来解码采集的信号。该项目的最终目标是拥有一个能够定量每个样品2，500个核酸靶标的系统，以比市售系统更好的灵敏度检测具有单碱基对识别的靶标，并以每分钟500个颗粒的速度扫描5，000个颗粒。该项目与公共卫生的相关性是为基因组医学开发一种变革性技术，从疾病诊断到药物发现。公共卫生相关性：该项目将开发一种新技术，可用于同时检测溶液中的数千种生物分子。这项新技术将在疾病诊断/治疗、提高输血安全性的血型定型和药物开发中找到潜在用途。