BIOASSAY WITH MAGNETIC PARTICLES IN FLOW

使用流动的磁性粒子进行生物测定

• 批准号: 7598415
• 负责人: MICHELLE Anna ESPY
• 金额: $ 2.1万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7598415
• 关键词: Binding; Biological Assay; Cell Cycle Regulation Pathway; Collection; Computer Retrieval of Information on Scientific Projects Database; DNA analysis; Detection; Diagnostic; Encapsulated; Flow Cytometry; Fluorescence; Funding; Goals; Grant; Health; Human; Incubated; Institution; Lasers; Magnetism; Measures; Methods; Microspheres; Molecular; Molecular Analysis; Molecular Target; Operative Surgical Procedures; Polymers; Proteomics; Quality of life; Range; Research; Research Personnel; Resources; Sorting - Cell Movement; Source; Squid; System; Technology; United States National Institutes of Health; Vesicle; base; commercialization; concept; detector; drug discovery; improved; instrument; instrumentation; magnetic field; novel; particle; therapeutic target; trafficking

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The goal of the project is to develop and demonstrate two related approaches to molecular analysis and separation that employ flow based analytical instrumentation and magnetic microsphere technology: a magnetic flow spectrometer for separation, and a magnetic flow cytometer for identification. The flow spectrometer system will be unique in enabling highly parallel continuous flow biomolecular separations on a preparative scale, streamlining downstream analysis and revolutionizing our ability to identify potential diagnostic or therapeutic targets. The magnetic flow cytometer will combine a novel magnetic target-molecule tagging concept with fluorescence-based analyte detection. The instrumentation proposed will contribute significantly to a broad range of applications improving human health and quality of life including drug discovery, molecular targeting, DNA analysis, proteomics, and understanding the pathways of cell cycle regulation. We will validate the new instrument by conventional molecular analysis methods and apply it to the study of intracellular vesicle traffic. A product for commercialization is anticipated. Operation of the proposed instrument involves three steps. 1) Magnetically encoded microspheres are prepared by encapsulating strong ferromagnetic material with high remnant magnetization and coercivity, never before used for such applications, in polymer spheres. The distribution of microspheres can be sorted into different bins depending on their intrinsic magnetic moment by flowing through a chamber where a magnetic field gradient induces a force such that they are collected in different bins with narrow distributions of magnetic moment. Microspheres from each bin are chemically bound to target molecules so that each species of magnetic moment is bound to one unique kind of molecule. The collection of microspheres and associated target molecules are then mixed together and incubated with analytes. 2) The incubated collection of microspheres are flowed through a SQUID detector system which identifies the target molecule by measuring the magnetic moment of the microsphere to which it is attached. 3) The analytes will be chemically prepared with molecular groups that fluoresce when illuminated by a laser beam, indicating the target-analyte binding. Combining SQUIDs for target identification with laser diagnostics to assess binding provides an efficient, high throughput multiplexed bioassay method based on traditional flow cytometry.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 该项目的目标是开发和演示使用流动分析仪器和磁性微球技术的两种相关的分子分析和分离方法：用于分离的磁性流动光谱仪和用于鉴定的磁性流式细胞仪。流动光谱仪系统将在制备规模上实现高度并行的连续流动生物分子分离，简化下游分析，并彻底改变我们识别潜在诊断或治疗靶点的能力。磁性流式细胞仪将把一种新的磁性目标分子标记概念与基于荧光的分析物检测相结合。所提出的仪器将对改善人类健康和生活质量的广泛应用做出重大贡献，包括药物发现、分子靶向、DNA分析、蛋白质组学和了解细胞周期调节的途径。我们将通过传统的分子分析方法对新仪器进行验证，并将其应用于细胞内囊泡运输的研究。预计会有一款商业化的产品。 拟议文书的运作包括三个步骤。1)磁性编码微球是通过将具有高剩余磁化强度和矫顽力的强铁磁性材料包裹在聚合物球中而制备的，这是以前从未用于此类应用的。微球的分布可以通过流过磁场梯度感应力的腔来根据其固有磁矩被分类到不同的箱中，从而使它们被收集在具有窄磁矩分布的不同箱中。来自每个储存箱的微球被化学绑定到目标分子上，因此每种磁矩都绑定到一种独特的分子上。收集的微球和相关的目标分子然后混合在一起，与分析物孵化。2)孵化的微球集合流经SQUID探测器系统，该系统通过测量目标分子所附着的微球的磁矩来识别目标分子。3)分析物将通过化学方法制备，分子基团在激光照射下会发出荧光，表明目标与分析物的结合。结合用于靶标识别的SQUID和用于评估结合的激光诊断，提供了一种基于传统的流式细胞术的高效、高通量的多重生物检测方法。