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 目前，年龄是发达国家主要慢性病的第一大危险因素。 平均 81% 的美国成年人患有一种或多种慢性疾病。 美国人口年龄持续增长，我们将面临重大挑战 如果患有慢性病的成年人的比例保持不变，那么疾病负担现在就很好。 确定炎症、衰老和与年龄相关的疾病的发展之间存在联系 潜在生物机制的复杂网络，但这些机制和联系 造成这种情况的一个主要原因是缺乏衡量它们之间的工具。 具体而言，在大浓度范围内同时检测所有相关生物标志物。 衰老研究和临床诊断非常需要一种可以测量的工具 定量、可靠、同时地检测多种炎症相关生物标志物 浓度范围大。 有一种技术如果更加灵敏的话，将是该应用的理想选择。 Luminex xMAP 系统经过充分验证，可测量多达 500 个生物标志物 同时，但这些测定通常无法提供给定的完整表征 由于系统的灵敏度限制，根本不可能使用 Luminex 进行采样。 我们的产品是一种检测低丰度生物标志物的技术，我们称之为纳米结构。 “Plasmonic Fluor”可用于增强 Luminex 检测，显着提高灵敏度 当前测定的局限性 确定了当前 Luminex 测定的灵敏度极限， 部分取决于目前用于检测生物标志物的报告荧光团的亮度。 使用的荧光团是 R-藻红蛋白 (PE)，这是一种荧光蛋白，是最亮的选择之一 我们有初步数据表明我们的等离激元荧光灯至少亮 50 倍。 比 PE 更容易被替代以显着改善 Luminex 检测，而不需要 检测工作流程或读出硬件的更改。 在这个项目中，我们的目标是：1) 优化我们的等离子体荧光以增强 Luminex 检测；2) 证明等离子体荧光增强的 Luminex 能够测量生物标志物 我们认为，其浓度比当前的 Luminex 系统低几个数量级。 Plasmonic Fluor 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 检测将成为事实上的标准工具，用于阐明 炎症、衰老和疾病之间的机制联系。