Plasmonic Detection of Free-Bilirubin in Neonates

新生儿游离胆红素的等离子体检测

• 批准号: 7313027
• 负责人: Christopher Darren Geddes
• 金额: $ 17.77万
• 依托单位: UNIVERSITY OF MD BIOTECHNOLOGY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7313027
• 关键词: Adult; Applications Grants; Area; Award; Back; Bilirubin; Binding; Biological; Biological Assay; Biomedical Engineering; Biotechnology; Blood; Blood - brain barrier anatomy; Calibration; Chemicals; Clinical; Clinical Research; Clinical Trials; Colloids; Dependence; Detection; Diffuse; Discipline; Enzyme-Linked Immunosorbent Assay; Face; Film; Fluorescence; Gestational Age; Glass; Gold; Health; Health Sciences; Housing; Human; Icterus; Island; Journals; Label; Laboratories; Link; Localized; Measurement; Measures; Metals; Methods; Molecular Weight; Nanostructures; Neonatal; Neurologic; Operative Surgical Procedures; Peer Review; Photons; Plastics; Polymerase Chain Reaction; Polymers; Premature Infant; Process; Publications; Purpose; RNA; Research; Research Project Grants; Risk; Sampling; Screening procedure; Serum; Serum Albumin; Silicon Dioxide; Silver; Slide; Staging; Standards of Weights and Measures; Surface; System; Technology; Testing; Thick; Thinking; Time; Today; Toxic effect; Translating; United States Food and Drug Administration; United States National Institutes of Health; Universities; Week; Whole Blood; base; copolymer; design; detector; fluorophore; hydrophilicity; improved; instrument; multidisciplinary; nanoparticle; neonate; nervous system disorder; new technology; novel; postnatal; quantum

DESCRIPTION (provided by applicant): Over the last couple of years our laboratories have developed a new fluorescence based phenomenon, which is now having profound implications in clinical biotechnology. This new technology has led to the recent publication of over 50 peer reviewed articles, one edited hard bound volume and the launch of a new peer reviewed Springer Journal, entitled Plasmonics. This new phenomenon relies on the interactions of fluorophores with metallic nanoparticles and can increase the quantum yield of fluorescing species, provide spatially localized excitation and can even substantially improve fluorophore photostability. Our new technology clearly demonstrates that these novel effects can result in up to a million-fold more photons per fluorophore, which when applied to assay sensing platforms, provides for quick and simple high sensitivity assays. In this regard, under a recently completed NIH award to the PI, we were able to develop an enhanced assay platform for RNA target detection which has the sensitivity of PCR and ELISA, but most profoundly, without any amplification steps. In this R21 proposal we have assembled a multidisciplinary team of Spectroscopists, Biostatisticians and Clinicians alike, to develop a high sensitivity assay for the detection of free unbound Bilirubin in neonatal serum, an area of intense research and high clinical importance due to its ability to cross the blood brain barrier and participate in various neurological disorders. At present, tests for free unbound bilirubin are arduous, time consuming, and no approach has to date been standardized. The Metal-Enhanced Fluorescence (MEF) free bilirubin assay described here, offers high sensitivity detection, simple operation, can be made disposable and has the potential to even be standardized in laboratories around the world. The MEF assay functions as follows: Human Serum Albumin (HSA), which binds and transports serum free and bound bilirubin respectively, is immobilized in a hydrophilic copolymer, itself immobilized onto a glass slide. When serum, containing serum albumin predominantly bound bilirubin (> 99 %) is washed across the assay surface, free bilirubin diffuses into the polymer, binds to the embedded HSA and becomes intrinsically highly fluorescent due to its locality to surface silver nanostructures. We intend to optimize and determine the sensitivity of the assay using both laboratory and clinical samples and most importantly, statistically compare our results with the "UB analyzer", one of only 3 instruments in the USA designed to measure free bilirubin. This proposed research aims to develop a highly novel sensing platform for whole blood free Bilirubin detection in neonates. No commercial instrument is available in the US today. Given that free bilirubin has been linked to later stage neurological disorders, then the technology underpinned by this proposal goes some significant ways to safeguard both neonatal and in-turn adult health.

描述（由申请人提供）：在过去的几年里，我们的实验室已经开发出一种新的基于荧光的现象，该现象现在在临床生物技术中具有深远的影响。这项新技术导致最近发表了50多篇同行评议的文章，一本编辑过的精装书，并推出了一本新的同行评议的Springer期刊，名为Plasmonics。这种新现象依赖于荧光团与金属纳米颗粒的相互作用，并且可以增加荧光物质的量子产率，提供空间局部激发，甚至可以显著改善荧光团的光稳定性。我们的新技术清楚地表明，这些新效应可以使每个荧光团产生高达一百万倍的光子，当应用于检测传感平台时，可以提供快速简单的高灵敏度检测。在这方面，根据最近完成的NIH对PI的奖励，我们能够开发用于RNA靶检测的增强型测定平台，其具有PCR和ELISA的灵敏度，但最重要的是，没有任何扩增步骤。在这项R21提案中，我们组建了一个由光谱学家、生物统计学家和临床医生组成的多学科团队，以开发一种用于检测新生儿血清中游离未结合胆红素的高灵敏度检测方法，这是一个密集研究和高度临床重要性的领域，因为它能够穿过血脑屏障并参与各种神经系统疾病。目前，游离未结合胆红素的测试是艰巨的，耗时的，并且迄今为止没有标准化的方法。本文所述的无金属增强荧光（MEF）胆红素测定提供了高灵敏度检测、操作简单、可一次性使用，甚至有可能在世界各地的实验室中标准化。MEF测定的功能如下：将分别结合和转运血清游离胆红素和结合胆红素的人血清白蛋白（HSA）固定在亲水性共聚物中，该共聚物本身固定在载玻片上。当含有主要结合胆红素（> 99%）的血清白蛋白的血清被洗涤穿过测定表面时，游离胆红素扩散到聚合物中，结合到包埋的HSA，并且由于其对表面银纳米结构的位置而变得固有地高度荧光。我们打算使用实验室和临床样本优化和确定测定的灵敏度，最重要的是，将我们的结果与“UB分析仪”进行统计学比较，UB分析仪是美国仅有的3种用于测量游离胆红素的仪器之一。本研究旨在开发一种用于新生儿全血游离胆红素检测的高度新颖的传感平台。目前在美国还没有商业仪器。鉴于游离胆红素与后期神经系统疾病有关，因此该提案所支持的技术在保护新生儿和成人健康方面发挥了重要作用。