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）：多路复用筛查是一种在药物发现，基因分型，医学诊断和血液键入等应用中发现广泛使用的工具，并在“个性化”药物的新兴领域中至关重要。这两种可商购的筛选技术提供了测量分析物（即平面微阵列）或高样品吞吐量（即基于珠子的系统）的高“密度”，但并非两者兼而有之。该应用程序提出了基于多功能编码粒子的新筛选技术的基本开发，该粒子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，其一半写成的打孔代码条形码和另一个靶标的条纹，可用于同时量化单个生物样品中的靶标，其编码能力超过一百万。在建立概念验证的演示中，假设（1）（1）增加水凝胶结构中的孔的大小将使靶标在整个粒子中结合，从而增加每个测定的灵敏度，同时减少所需的孵育时间； （2）可以调整杂交条件以实现与现有技术的性能竞争； （3）基于微流体和光电层流技术的流通系统可用于快速扫描颗粒（即读取代码并量化目标）。该项目的具体目的是：（1）通过探索化学变化和加工条件来产生足够多的多孔且机械稳健的颗粒来增强颗粒合成。将通过显微镜检查颗粒，并用FITC偶联的右旋体探测。 （2）优化DNA杂交测定的物理和化学条件，以最大程度地提高灵敏度，特异性和可重复性。 （3）开发一种基于微流体流的系统，用于快速扫描，该系统集成了流动的微流体设备，光电倍增器荧光检测和软件来解码所获得的信号。该项目的最终目标是拥有一个能够量化每个样品2,500个核酸靶标的系统，以比市售系统更好的灵敏度检测单碱基对歧视的目标，并以每分钟500个颗粒的速率扫描5,000个颗粒。该项目与公共卫生的相关性是开发用于基因组医学的变革性技术，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，该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/治疗中发现潜在用途，以提高输血和药物发育的安全性。