A High Sensitivity Multiplexed Immunoassay for Aging-Related Inflammation Biomarker Quantification

用于衰老相关炎症生物标志物定量的高灵敏度多重免疫测定

• 批准号: 9909594
• 负责人: Scott L Crick
• 金额: $ 29.96万
• 依托单位: AURAGENT BIOSCIENCE, LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9909594
• 关键词: Adult; Age; Aging; Antibodies; Bar Codes; Binding; Biological; Biological Assay; Biological Markers; Biotin; Bovine Serum Albumin; Characteristics; Chronic Disease; Data; Detection; Development; Disease; Dyes; Enzyme-Linked Immunosorbent Assay; Fluorescence; Gold; Home environment; Immobilization; Immunoassay; Inflammation; Label; Link; Manufacturer Name; Measurement; Measures; Metals; Microspheres; Nanostructures; Peer Review; Performance; Photobleaching; Phycoerythrin; Population; Process; Proteins; Publications; Reagent; Reporter; Reproducibility; Research; Research Personnel; Rhodamine; Risk Factors; Role; Sampling; Silver; Streptavidin; Structure; System; Technology; Testing; Thick; United States; Universities; Washington; Work; age related; base; burden of illness; clinical diagnostics; cost effective; density; design; experience; fluorophore; improved; inflammatory marker; nanorod; plasmonics; prototype; scaffold; specific biomarkers; tool

Abstract Age is the number one risk factor for the major chronic diseases in the developed world. Currently, 81% of adults in the United States are afflicted with one or more chronic diseases. As the average age of the population of the United States continues to grow, we will be faced with a significant disease burden if the percentage of adults with chronic disease remains the same. It is now well established that inflammation, aging, and the development of age-related diseases are linked by a complicated network of underlying biological mechanisms, but these mechanisms and the links between them are not well understood. A major reason for this is the lack of a tool to measure the all of the relevant biomarkers simultaneously over a large range of concentrations. Specifically, there is major need in aging research and clinical diagnostics for a tool which can measure multiple inflammation-related biomarkers quantitatively, reliably, and simultaneously over a large range of concentrations. There is a technology that would be ideal for this application if it were simply more sensitive. The Luminex xMAP system is well-validated and allows measurement of up to 500 biomarkers simultaneously, but these assays often fail to provide a complete characterization of a given sample due to sensitivity limitations of the system. It is simply not possible to use the Luminex technology to detect low abundance biomarkers. Our product, a nanoconstruct which we call a “Plasmonic Fluor”, can be used to enhance Luminex assays to significantly improve the sensitivity limitations of the current assays. The sensitivity limit of the current Luminex assays is determined, in part, by the brightness of the reporter fluorophore used in detecting biomarkers. The currently used fluorophore is R-phycoerythrin (PE), a fluorescent protein that is one of the brightest options available. We have preliminary data suggesting our Plasmonic Fluors are at least 50X brighter than PE, and can easily be substituted to significantly improve Luminex assays without requiring changes to the assay workflow or readout hardware. In this project, we aim to: 1) optimize our Plasmonic Fluor for enhancing Luminex assays; and 2) demonstrate that Plasmonic Fluor-enhanced Luminex is capable of measuring biomarkers at concentrations that are orders of magnitude lower than the current Luminex system. We believe the Plasmonic Fluor will completely replace PE as the standard reporter fluorophore, and Plasmonic Fluor-enhanced Luminex assays will become the de facto standard tool for elucidating the mechanistic linkages between inflammation, aging, and disease.

抽象的 年龄是发达国家主要慢性疾病的第一危险因素。现在， 美国有81％的成年人患有一种或多种慢性疾病。作为平均水平 美国人口的年龄不断增长，我们将面临重要的 疾病伯宁如果患有慢性疾病的成年人的百分比保持不变。现在很好 确定感染，衰老和与年龄有关疾病的发展是由 基本生物学机制的复杂网络，但是这些机制和联系 他们之间的理解不太了解。造成这种情况的主要原因是缺乏测量工具 所有相关的生物标志物都只是在众多浓度范围内。具体来说， 对可以测量的工具的衰老研究和临床诊断非常需要 多种炎症相关的生物标志物数量，可靠地且简单地结束 浓度范围很大。 如果它简单地敏感，就有一项技术对于此应用程序是理想的选择。这 Luminex Xmap系统经过验证，可允许多达500个生物标志物测量 同样，但是这些测定通常无法提供给定的完整表征 由于系统的敏感性限制而引起的样本。根本无法使用Luminex 检测低抽象生物标志物的技术。我们的产品，一种我们称为的纳米结构 “等离荧光”可用于增强Luminex分析以显着提高灵敏度 当前测定法的局限性。确定当前Luminex分析的灵敏度极限， 在某种程度上，通过用于检测生物标志物的记者荧光团的亮度。当前 使用的荧光团是R-Phycoerythrin（PE），这是一种荧光蛋白，是最亮的选择之一 可用的。我们有初步数据表明我们的等离子荧光至少更明亮50倍 比PE，可以轻松替代以显着改善Luminex分析而无需 更改测定工作流程或读数硬件。 在这个项目中，我们的目标是：1）优化我们的等离子荧光以增强Luminex分析；和2） 证明等离激氟增强的Luminex能够测量生物标志物 浓度比当前的Luminex系统低。我们相信 等离子体荧光将完全取代PE作为标准报告荧光团，并且 等离子体荧光增强的Luminex分析将成为阐明的事实上的标准工具 炎症，衰老和疾病之间的机械联系。