Nanocaged Metal Tags in Massively Multiplexed Leukemia Bioassay and Beyond

大规模多重白血病生物测定及其他领域中的纳米笼金属标签

• 批准号: 8504748
• 负责人: Janice Paige Buchanan
• 金额: $ 12.47万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8504748
• 关键词: Acute Myelocytic Leukemia; Acute leukemia; Address; Advanced Development; Affinity; Alkenes; Antibodies; Antigens; Binding; Biological Assay; Biological Markers; Carbon; Cells; Characteristics; Classification; Colorado; Coupled; Detection; Diagnosis; Disease; Elements; Feedback; Flow Cytometry; Fullerenes; Gene Expression Profiling; Gene Proteins; Genes; Goals; Growth Factor; Healthcare; Human; Immunoassay; Investigation; Life; Link; Mass Spectrum Analysis; Measurement; Medical; Metals; Microspheres; Mississippi; Oligonucleotides; Patients; Plasma; Polymers; Preparation; Proteins; Relative (related person); Reporter; Robotics; Sampling; Serum; Surface; System; Techniques; Technology; Testing; Time; Transcript; Universities; Validation; Work; analytical method; base; chemical synthesis; chemokine; cytokine; functional group; instrumentation; leukemia; light scattering; mass spectrometer; monomer; nanocage; novel; outcome forecast; polymerization; receptor; small molecule

DESCRIPTION (provided by applicant): Characterization of an acute leukemia (AML) cell requires measurement of biomarkers such as proteins, genes and small molecules. An unambiguous biomarker signature cannot be obtained by determining only a few proteins. A quantitative, massively multiplexed bioassay (MMB) for a constellation of proteins, small molecules, and gene transcripts would be a significant medical breakthrough. "Personalized health care", unambiguous disease identification, and patient- specific diagnosis/prognosis will be enabled by the simultaneous quantitative determination of many biomarkers in a patient's sample. We will develop a new, robust, multimetallic tagging system based on functionalized polymer microbeads (PMBs) analyzed by the proven, high-sensitivity, time-resolved, and element-specific detection technique of flow-cytometry inductively- coupled-plasma mass spectrometry (FC-ICP-MS). Our interdisciplinary group of recognized experts will address this significant challenge. Prof. S. Stevenson (Univ. of Southern Mississippi, USM) will optimize the synthesis of 7 different M3N@C80 endometallofullerenes (EMFs) with interference-free metals that are not endogenous in biomedical samples. Prof. S. H. Strauss and Dr. O. V. Boltalina (Colorado State Univ., CSU) will optimize the synthesis of chemical derivatives of the EMFs, determine the optimum type and number of functional groups to achieve uniformity. Preferred derivatives will be sent to Prof. J. P. Phillips (USM), who will attach polymerizable groups to EMF derivatives, optimize preparation of narrow-size-distribution PMBs containing many specific mixtures of the seven metals (all of which will be permanently sequestered in their carbon nanocages and therefore cannot leach out over time). The Univ. of Toronto (UofT) group, led by Prof. V. I. Baranov, will develop purpose-specific analytical methods that combine robotic sample introduction and FC-ICP-MS instrumentation to allow massively multiplexed detection and classification of thousands of multimetallic encoded beads with element-tagged reporter affinity molecules. They will estimate the tagging multiplexity and show the integrity and virtually infinite shelf-life of the tagged PMBs, and that massive multiplexing with thousands of differently tagged PMBs is possible when this technology is reduced to practice. Dr. O. I. Ornatsky (UofT) will perform analyte selection, testing and validation of the encoded beads, immunoassays, and oligonucleotide hybridization, and will covalently link a range of antibodies (Abs) to the surfaces of the tagged PMBs, which will be tested in sandwich assays with secondary Abs linked to a reporter tag. We will prove that our tagging system has the potential to be used for massively multiplexed bioassays in which thousands of antigens, gene transcripts, and small-molecule cell markers for many diseases and conditions are determined simultaneously in a single sample. We will test our massively multiplexed bioanalytical platform on acute myeloid leukemia (AML) cells. We will show the advantages of FC-ICP-MS analysis of multi-metal-tagged PMBs coated with capture Abs against cytokines, chemokines, growth factors, and soluble receptors present in human serum.

描述（由申请人提供）：急性白血病（AML）细胞的表征需要测量生物标志物，例如蛋白质，基因和小分子。明确的生物标志物签名无法通过仅确定几种蛋白质来获得。用于蛋白质，小分子和基因转录本的定量，大量多重生物测定（MMB）将是一个显着的医学突破。 “个性化医疗保健”，明确的疾病鉴定以及患者特定的诊断/预后将通过同时定量确定患者样本中的许多生物标志物。我们将根据功能化的聚合物微头（PMB）开发一种新的，可靠的多功能标记系统，该系统通过经过验证的，高敏，时间分辨和元素特异性检测技术分析的功能化聚合物微粒（PMB），对流动感应式感应型辅助 - 偶发性 - 倍增 - 倍膜质谱法（FC-ICP-MS）。我们公认的专家跨学科小组将应对这一重大挑战。 S. Stevenson教授（USM密西西比州南部）将优化7种不同的M3N@C80@c80内膜甲氟列烯（EMFS），其无干扰金属在生物医学样品中不是内源性的。 S. H. Strauss教授和O. V. Boltalina博士（Colorado State Univ。，CSU）将优化EMF的化学衍生物的合成，确定实现均匀性的最佳类型和数量。首选的衍生物将被发送给J. P. Phillips教授（USM），后者将将可聚合基团附加到EMF衍生物上，优化制备狭窄的大小分布PMB，其中包含七种特定的七种特定混合物（所有这些都将在其碳纳米含量中永久性地进行固定，因此不能随着时间的推移渗出）。大学。由V. I. Baranov教授领导的多伦多（UOFT）组将开发特定于特定的分析方法，这些方法将机器人样品引入和FC-ICP-MS仪器结合起来，以允许大量多路复用检测和成千上万个多属金属编码珠的分类和分类。他们将估计标记的多重性，并显示标记的PMB的完整性和几乎无限的保质期，并且当该技术降低到练习时，可以使用数千种具有不同标记的PMB的大量多路复用。 O. I. Ornatsky博士（UOFT）将对编码珠，免疫测定和寡核苷酸杂交进行选择，测试和验证，并将共价将一系列抗体（ABS）连接到标记PMB的表面，并将其在夹心链接中进行了测试，该抗体将在sandwich link link sandare ables link sandare linked tag链接中进行测试。我们将证明我们的标记系统有可能用于大规模多重生物测定，其中数千种抗原，基因转录本和用于许多疾病的小分子细胞标记在单个样本中同时确定。我们将在急性髓样白血病（AML）细胞上测试大量多重生物分析平台。我们将展示与捕获ABS相对于人血清中存在的细胞因子，趋化因子，生长因子和可溶性受体的FC-ICP-MS分析的优势。

项目成果

Structure of 7,9,12,15,18,20,39,24,45,57-C60(CF3)10(1,2:3,4-O)2. The first regiospecific diepoxidation of a fullerene derivative.

• 发表时间: 2013-11
• 期刊: Acta chimica Slovenica
• 影响因子: 1.2
• 作者: James B. Whitaker;N. Shustova;S. Strauss;B. V

Perfluoroalkylfullerenes.

• DOI: 10.1021/cr5002595
• 发表时间: 2015-01-28
• 期刊: CHEMICAL REVIEWS
• 影响因子: 62.1
• 作者: Boltalina, Olga V.;Popov, Alexey A.;Kuvychko, Igor V.;Shustova, Natalia B.;Strauss, Steven H.
• 通讯作者: Strauss, Steven H.