Bioassay with magnetic particles in flow

使用流动中的磁性颗粒进行生物测定

• 批准号: 7463624
• 负责人: MICHELLE Anna ESPY
• 金额: $ 39.69万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-15 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7463624
• 关键词: Affect; Binding; Biological; Biological Assay; Cell Cycle Regulation Pathway; Collection; Coulter counter; DNA analysis; Data; Detection; Devices; Diagnostic; Encapsulated; Flow Cytometry; Fluorescence; Force of Gravity; Genomics; Goals; Health; Human; Incubated; Individual; Lasers; Ligands; Liquid substance; Magnetism; Measurement; Measures; Methods; Microscopy; Microspheres; Modeling; Molecular; Molecular Analysis; Molecular Target; Noise; Numbers; Operative Surgical Procedures; Optics; Performance; Phase; Polymers; Population; Preparation; Property; Proteomics; Quality of life; Range; Rate; Reproducibility; Resolution; Sampling; Sorting - Cell Movement; Squid; System; Techniques; Technology; Time; Validation; Vesicle; Viscosity; base; commercialization; concept; density; design; detector; drug discovery; improved; instrument; instrumentation; magnetic field; novel; novel strategies; particle; physical property; size; therapeutic target; trafficking

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.

该项目的目标是开发和演示两种相关的分子分析方法和 采用基于流动的分析仪器和磁性微球技术的分离： 磁流谱仪用于分离，磁流式细胞仪用于鉴定。流量 光谱仪系统将在实现高度并行连续流生物分子分离方面具有独特性。 制备规模，简化下游分析并彻底改变我们识别潜力的能力 诊断或治疗目标。磁流式细胞仪将结合一种新颖的磁性靶标—— 分子标记概念与基于荧光的分析物检测。拟议的仪器将 为改善人类健康和生活质量的广泛应用做出了重大贡献，包括 药物发现、分子靶向、DNA 分析、蛋白质组学以及了解细胞周期途径 规定。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 由封装具有高剩磁和矫顽力的强铁磁材料制成， 以前从未在聚合物球中用于此类应用。可对微球的分布进行排序 通过流过一个腔室，根据其固有磁矩进入不同的容器，其中 磁场梯度会产生一种力，使它们被收集到分布较窄的不同容器中 的磁矩。每个容器中的微球都与目标分子进行化学结合，以便每个容器 磁矩种类与一种独特的分子结合。微球的收集和 然后将相关的目标分子混合在一起并与分析物一起孵育。 2) 孵化 微球的集合流过 SQUID 检测器系统，该系统识别目标分子 通过测量其所附着的微球的磁矩。 3) 分析物将是 用化学方法制备的分子基团在激光束照射下会发出荧光，表明 目标-分析物结合。将用于目标识别的 SQUID 与激光诊断相结合以评估结合 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。