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.

摘要 在发达国家，年龄是主要慢性病的头号危险因素。目前， 在美国，81%的成年人患有一种或多种慢性疾病。随着平均 美国人口年龄持续增长，我们将面临重大挑战 如果成年人慢性病患者的比例保持不变，疾病负担将减少。现在好了 确定炎症，衰老和年龄相关疾病的发展与以下因素有关： 一个复杂的潜在生物机制网络，但这些机制和联系 他们之间的关系并不清楚。一个主要原因是缺乏一个工具来衡量 所有相关的生物标志物同时在一个大的浓度范围。具体地说， 在老化研究和临床诊断中主要需要一种工具 多种炎症相关的生物标志物定量、可靠、同时超过 浓度范围很大。 有一种技术，将是理想的这种应用程序，如果它只是更敏感。的 Luminex xMAP系统经过充分验证，可测量多达500种生物标志物 但是这些测定通常不能提供给定的细胞的完整表征。 由于系统的灵敏度限制，使用Luminex是不可能的 低丰度生物标志物检测技术。我们的产品，一种纳米结构，我们称之为 “Plasmonic Fluor”可用于增强Luminex测定，以显著提高灵敏度 当前检测的局限性。确定当前Luminex测定的灵敏度限值， 部分取决于用于检测生物标志物的报道荧光团的亮度。当前 使用的荧光团是R-藻红蛋白（PE），一种荧光蛋白，是最亮的选择之一 available.我们有初步的数据表明我们的等离子体荧光灯至少亮50倍 并且可以容易地替代以显著改善Luminex测定，而不需要 分析工作流程或读出硬件的变更。 在这个项目中，我们的目标是：1）优化我们的Plasmonic Fluor，以增强Luminex检测; 2） 证明Plasmonic Fluor-enhanced Luminex能够测量生物标志物， 浓度的数量级低于目前的Luminex系统。我们认为 Plasmonic Fluor将完全取代PE作为标准报告荧光团， Plasmonic Fluor-enhanced Luminex检测将成为阐明 炎症、衰老和疾病之间的机械联系。