BIOASSAY WITH MAGNETIC PARTICLES IN FLOW

使用流动的磁性粒子进行生物测定

• 批准号: 8361760
• 负责人: MICHELLE Anna ESPY
• 金额: $ 1.12万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361760
• 关键词: Binding; Biological Assay; Cell Cycle Regulation Pathway; Collection; DNA analysis; Detection; Diagnostic; Encapsulated; Flow Cytometry; Fluorescence; Funding; Goals; Grant; Health; Human; Incubated; Lasers; Magnetism; Measures; Methods; Microspheres; Molecular; Molecular Analysis; Molecular Target; National Center for Research Resources; Polymers; Principal Investigator; Proteomics; Quality of life; Research; Research Infrastructure; Resources; Sorting - Cell Movement; Source; System; Technology; United States National Institutes of Health; Vesicle; base; commercialization; cost; detector; drug discovery; improved; instrument; instrumentation; magnetic field; novel; operation; particle; therapeutic target; trafficking

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 该项目的目标是开发和演示两种相关的分子分析和分离方法，它们采用基于流的分析仪器和磁性微球技术：用于分离的磁流光谱仪和用于识别的磁流式细胞仪。流式光谱仪系统的独特之处在于能够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。磁流式细胞仪将新颖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。 所提出的仪器将为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控途径。我们将通过常规分子分析方法验证新仪器，并将其应用于细胞内囊泡运输的研究。 预计将有产品商业化。 拟议仪器的操作涉及三个步骤。 1) 磁编码微球是通过将具有高剩磁和矫顽力的强铁磁材料封装在聚合物球中而制备的，以前从未用于此类应用。微球的分布可以根据其固有磁矩通过流过一个室而被分类到不同的箱中，在该室中，磁场梯度感生力，使得它们被收集在具有窄磁矩分布的不同箱中。每个容器中的微球都与目标分子进行化学结合，以便每种磁矩与一种独特的分子结合。然后将微球和相关目标分子的集合混合在一起并与分析物一起孵育。 2) 孵育后的微球体流过 SQUID 检测器系统，该系统通过测量附着的微球体的磁矩来识别目标分子。 3) 分析物将用在激光束照射时发出荧光的分子基团进行化学制备，表明目标分析物的结合。将用于目标识别的 SQUID 与用于评估结合的激光诊断相结合，提供了一种基于传统流式细胞术的高效、高通量多重生物测定方法。