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。仅通过确定少数蛋白质无法获得明确的生物标志物特征。针对蛋白质、小分子和基因转录本的定量、大规模多重生物测定（MMB）将是一项重大的医学突破。通过同时定量测定患者样本中的许多生物标志物，将能够实现“个性化医疗保健”、明确的疾病识别和患者特异性诊断/预后。我们将开发一种基于功能化聚合物微珠 (PMB) 的新型、稳健的多金属标记系统，并通过经过验证的高灵敏度、时间分辨和元素特异性检测技术流式细胞术电感耦合等离子体质谱 (FC-ICP-MS) 进行分析。我们的跨学科专家小组将解决这一重大挑战。 S. Stevenson 教授（南密西西比大学，USM）将利用生物医学样品中非内源性的无干扰金属来优化 7 种不同的 M3N@C80 内金属富勒烯 (EMF) 的合成。 S. H. Strauss 教授和 O. V. Boltalina 博士（科罗拉多州立大学，科罗拉多州立大学）将优化 EMF 化学衍生物的合成，确定最佳类型和官能团数量以实现均匀性。优选的衍生物将发送给 J. P. Phillips 教授（USM），他将把可聚合基团附加到 EMF 衍生物上，优化包含七种金属的许多特定混合物的窄尺寸分布 PMB 的制备（所有这些金属都将永久隔离在碳纳米笼中，因此不会随着时间的推移而浸出）。大学。由 V. I. Baranov 教授领导的多伦多大学 (UofT) 团队将开发特定用途的分析方法，将机器人进样和 FC-ICP-MS 仪器相结合，以实现对数千个带有元素标记报告亲和分子的多金属编码珠进行大规模多重检测和分类。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，并且当该技术付诸实践时，数千个不同标记的 PMB 的大规模多重分析是可能的。 O. I. Ornatsky 博士 (UofT) 将进行分析物选择、编码珠的测试和验证、免疫测定和寡核苷酸杂交，并将一系列抗体 (Ab) 共价连接到标记的 PMB 表面，这些抗体将在夹心测定中进行测试，二级 Ab 与报告标签相连。我们将证明我们的标记系统有潜力用于大规模多重生物测定，其中在单个样品中同时测定许多疾病和病症的数千种抗原、基因转录本和小分子细胞标记。我们将在急性髓系白血病 (AML) 细胞上测试我们的大规模多重生物分析平台。我们将展示 FC-ICP-MS 分析涂有针对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捕获抗体的多金属标记 PMB 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.

全氟烷基富勒烯。

• 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.