Development of (Prototype) Bead Array Flow Cytometer with Mass Spectrometer Detec

开发带有质谱仪 Detec 的珠阵列流式细胞仪（原型）

• 批准号: 7586381
• 负责人: Scott David Tanner
• 金额: $ 18.94万
• 依托单位: UNIVERSITY OF TORONTO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7586381
• 关键词: Advanced Development; Algorithms; Architecture; Area; Benchmarking; Binding; Biological Assay; Biological Markers; Caring; Cells; Chemistry; Clinics and Hospitals; Code; Commit; Communities; Computer software; Computers and Advanced Instrumentation; Cost-Benefit Analysis; Coupled; Data; Detection; Development; Device or Instrument Development; Devices; Diagnostic; Disease; Early Diagnosis; Effectiveness; Elements; Engineering; Evaluation; Financial cost; Flow Cytometry; Fluorescent Dyes; Fluorochrome; Gene Proteins; Genes; Genomics; Goals; Healthcare; Human; Immunoassay; Individual; Investigation; Ions; Label; Laboratories; Latex Bead; Mass Spectrum Analysis; Metals; Methodology; Modification; Molecular; Molecular Profiling; Oligonucleotides; Operative Surgical Procedures; Optics; Output; Patients; Phase; Physicians; Plasma; Protein Array; Proteomics; Reaction; Reagent; Research; Research Personnel; Resolution; Sampling; Scanning; Software Design; Solutions; Speed; Surface; System; Technology; Time; Transcript; Translations; Treatment Efficacy; Validation; analytical method; analytical tool; base; clinical application; cost; design; detector; experience; improved; innovation; instrument; instrumentation; mass spectrometer; miniaturize; new technology; novel; particle; prognostic; prototype; research and development; small molecule; stable isotope; technology development; tool; user-friendly

DESCRIPTION (provided by applicant): The unambiguous identification of a rare, diseased cell for the purpose of early diagnosis requires the quantitative determination of many biomarkers simultaneously in individual cells at high throughput. A similar requirement for multi-parameter assay is shared by genomic and proteomic researchers, who need to determine simultaneously the presence and quantity of the many genes, proteins and small molecules involved in the translation of gene to cell activity. We propose a novel solution for a broad spectrum of bio-analytical challenges. The new technology takes advantage of the high resolution and sensitivity of inductively coupled plasma mass spectrometry (ICP-MS), combined with the many available stable isotopes of elements of the periodic table, to simultaneously determine many proteins and gene transcripts in samples through the quantification of stable isotope tags bound to a wide variety of bioaffinity molecules. Beads are an attractive option for supporting surface chemistries for immunoassays and oligonucleotide hybridization assays. One of the advantages of beads is the ability to increase the reaction surface area per volume of the reaction mixture, which provides a reliable means of increasing the capacity and dynamic range potential of an assay, as well as miniaturizing the reaction. We expect that our technology will be able to recognize many thousands of distinguishable analytical beads created by the incorporation of various concentrations and ratios of metal ions. We hope that this integrated massively multiplexed technology based on our prototype flow cytometer-mass spectrometer (FC-MS) and reagent support/encoding system will simplify and enhance the diagnostic, prognostic and therapeutic efficacy available to physicians and their patients, increasing the effectiveness of healthcare while substantially reducing the human and financial costs of treatment. The objective of this application is the development of analytical instrumentation, an advanced element (metal) encoding system and methodology for rapid recognition of functionalized encoded beads - the bead array mass spectrometer (BA-MS). The combination with an elemental detector will provide researchers and clinicians with massively multiplexed analytical capabilities. The focus of this application is on the development of an individual (cytometric) bead analysis at high throughput (employing the FC-MS invented, developed and built in our lab). FC-MS continues to undergo extensive development in our lab and is not yet available elsewhere. Our Specific Aims include: 1) development of these technologies to the engineering beta-level, suitable for placement in laboratories for routine use and evaluation. These beta instruments will be made available within 12 months of the completion of the proposed effort. 2) development of a statistically significant unambiguous encoding system with massive variability (>105 individual beads) and unsupervised algorithms to recognize encoding in a rapid and dependable manner. 3) development of demonstrative bio-analytical methods using the technology in a multiplexed format and validation against existing fluorochrome bead-based technologies. Our team has long experience in research and pre-commercial analytical instrument development, is completely committed to the goals and is poised to advance rapidly.

描述（由申请人提供）：为了早期诊断的目的，明确鉴定罕见的患病细胞需要以高通量同时定量测定单个细胞中的许多生物标志物。基因组学和蛋白质组学研究人员对多参数测定也有类似的要求，他们需要同时确定参与基因到细胞活性翻译的许多基因、蛋白质和小分子的存在和数量。我们提出了一种新的解决方案，以应对广泛的生物分析挑战。新技术利用电感耦合等离子体质谱（ICP-MS）的高分辨率和灵敏度，结合元素周期表中元素的许多可用稳定同位素，通过定量与各种生物亲和分子结合的稳定同位素标签，同时确定样品中的许多蛋白质和基因转录本。珠粒是用于支持免疫测定和寡核苷酸杂交测定的表面化学的有吸引力的选择。珠粒的优点之一是能够增加每体积反应混合物的反应表面积，这提供了增加测定的容量和动态范围潜力以及使反应稳定化的可靠手段。我们期望我们的技术能够识别通过掺入各种浓度和比例的金属离子而产生的数千种可区分的分析珠。我们希望这种基于我们的原型流式细胞仪-质谱仪（FC-MS）和试剂支持/编码系统的集成大规模多路复用技术将简化和增强医生及其患者的诊断，预后和治疗效果，提高医疗保健的有效性，同时大幅降低治疗的人力和财务成本。本申请的目的是开发分析仪器，先进的元素（金属）编码系统和方法，用于快速识别功能化编码珠-珠阵列质谱仪（BA-MS）。与元素探测器的结合将为研究人员和临床医生提供大规模多路复用的分析能力。本申请的重点是开发高通量的单个（细胞计数）珠粒分析（采用我们实验室发明、开发和构建的FC-MS）。FC-MS在我们的实验室继续进行广泛的开发，在其他地方还没有。我们的具体目标包括：1）将这些技术发展到工程测试水平，适合放置在实验室中进行常规使用和评估。这些测试工具将在拟议工作完成后12个月内提供。2)开发具有统计学显著性的明确编码系统，该系统具有巨大的可变性（>105个单个珠）和无监督算法，以快速可靠的方式识别编码。3)使用该技术以多路复用形式开发示范性生物分析方法，并针对现有的基于荧光染料珠的技术进行验证。我们的团队在研究和商业前分析仪器开发方面拥有长期经验，完全致力于实现目标，并准备迅速发展。