Plasmonic Detection of Free-Bilirubin in Neonates

项目概要/摘要： 在过去的几年里，我们的实验室开发了一种新的基于荧光的现象， 现在对临床生物技术有着深远的影响。这项新技术导致了最近 出版50多篇同行评审文章，一本编辑过的精装本，并推出一本新的同行评审论文集 Springer Journal的Plasmonics。这种新现象依赖于以下因素的相互作用： 荧光团与金属纳米颗粒的结合，并且可以增加荧光物质的量子产率， 空间局部激发，甚至可以显著改善荧光团光稳定性。我们的新 技术清楚地表明，这些新的效果可以导致高达百万倍以上的光子， 荧光团，其在应用于测定传感平台时提供快速和简单的高灵敏度 测定。在这方面，根据最近完成的NIH对PI的奖励，我们能够开发一种增强的 用于RNA靶标检测的分析平台，其具有PCR和ELISA的灵敏度，但最重要的是， 无需任何扩增步骤。在这个R21提案中，我们组建了一个多学科团队， 光谱学家、生物统计学家和临床医生等，开发一种高灵敏度的检测方法， 新生儿血清中游离未结合胆红素是一个密集的研究领域，由于其 能够穿过血脑屏障并参与各种神经系统疾病。目前，测试 游离未结合胆红素是费力、耗时的，并且迄今为止没有标准化的方法。的 这里描述的无金属增强荧光（MEF）胆红素测定提供高灵敏度检测， 操作简单，可以一次性使用，甚至有可能在实验室中标准化 世界各地MEF测定的功能如下：人血清白蛋白（HSA），其结合并 分别转运血清游离胆红素和结合胆红素，固定在亲水共聚物中，本身 固定在载玻片上。当血清中含有主要结合胆红素的血清白蛋白（> 99 %）被洗涤穿过测定表面，游离胆红素扩散到聚合物中，结合到包埋的HSA上， 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检测的灵敏度， 重要的是，将我们的结果与“UB分析仪”进行统计比较，UB分析仪是美国仅有的3种仪器之一 用于测量游离胆红素

Project summary/Abstract: 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.