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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

基本信息

批准号：
7777791
负责人：
Patrick S Doyle
金额：
$ 19.73万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7777791
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

项目摘要

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）：多重筛查是一种广泛应用于药物发现、基因分型、医学诊断和输血安全血型等应用的工具，并且在新兴的“个性化”医学领域至关重要。这两种市售筛选技术要么提供高“密度”的测量分析物（即平面微阵列），要么提供高样品通量（即基于微珠的系统），但不能同时提供两者。该应用提出了一种基于多功能编码颗粒的新筛选技术的根本发展，该技术可以提供微阵列的密度和基于珠子的系统的通量。初步结果表明，由海绵状水凝胶材料组成的颗粒，一半写有打孔码条形码，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本中的目标，编码能力超过百万级。在概念验证演示的基础上，假设 (1) 增加水凝胶结构中的孔尺寸将允许靶标结合在整个颗粒中，从而提高每次测定的灵敏度，同时减少所需的孵育时间； (2)可以调整杂交条件以实现与现有技术竞争的性能； (3) 基于微流体和光电倍增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，以生成足够多孔且机械坚固的颗粒。将通过显微镜检查颗粒并用 FITC 缀合的葡聚糖进行探测。 (2) 优化DNA杂交测定的物理和化学条件，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的快速扫描系统，该系统集成了流聚焦微流控装置、光电倍增管辅助荧光检测和解码所获取信号的软件。该项目的最终目标是拥有一个能够量化每个样本 2,500 个核酸目标的系统，以比市售系统更高的灵敏度检测具有单碱基对区分的目标，并以每分钟 500 个粒子的速度扫描 5,000 个粒子。该项目与公共卫生的相关性在于开发基因组医学的变革性技术，范围从疾病诊断到药物发现。公共健康相关性：该项目将开发一种新技术，可用于同时检测解决方案中的数千种生物分子。这项新技术将在疾病诊断/治疗、血型定型以提高输血安全和药物开发方面具有潜在用途。