BIOASSAY WITH MAGNETIC PARTICLES IN FLOW

使用流动的磁性粒子进行生物测定

• 批准号: 8169396
• 负责人: MICHELLE Anna ESPY
• 金额: $ 1.67万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169396
• 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; Polymers; Proteomics; Quality of life; Research; Research Personnel; Resources; Sorting - Cell Movement; Source; Squid; System; Technology; United States National Institutes of Health; Vesicle; base; commercialization; 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. 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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 该项目的目标是开发和展示两种相关的分子分析和分离方法，采用基于流动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鉴定的磁流式细胞仪。该流动光谱仪系统将在制备规模上实现高度平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方面是独一无二的。磁流式细胞仪将结合联合收割机一个新的磁性目标分子标记的概念与基于荧光的分析物检测。 所提出的仪器将大大有助于改善人类健康和生活质量的广泛应用，包括药物发现，分子靶向，DNA分析，蛋白质组学和理解细胞周期调控的途径。我们将通过传统的分子分析方法验证新仪器，并将其应用于胞内囊泡交通的研究。 预计将有一个商业化的产品。 拟议文书的运作涉及三个步骤。1)磁编码微球是通过将具有高剩余磁化强度和磁化率的强铁磁材料封装在聚合物球体中来制备的，以前从未用于此类应用。微球的分布可以根据它们的固有磁矩通过流过其中磁场梯度感应力的室而被分选到不同的仓中，使得它们被收集在具有窄磁矩分布的不同仓中。来自每个仓的微球与目标分子化学结合，使得每种磁矩与一种独特的分子结合。然后将微球和相关靶分子的集合混合在一起并与分析物一起孵育。2)使孵育的微球集合流过SQUID检测器系统，SQUID检测器系统通过测量其所附着的微球的磁矩来鉴定靶分子。3)分析物将用分子基团进行化学制备，当被激光束照射时，这些分子基团发出荧光，指示目标-分析物结合。将SQUID用于靶标识别与激光诊断结合以评估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。