Novel label-free electric immunosensing for detecting CVD markers

淡江大學機構典藏 > 理學院 > 化學學系暨研究所 > 學位論文 > Item 987654321/51834

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题名:	Novel label-free electric immunosensing for detecting CVD markers
其它题名:	新型無標定生醫免疫電學感測技術之開發
作者:	林志成;Lin, Chih-chen
贡献者:	淡江大學化學學系博士班李世元;Lee, Adam Shih-yuan
关键词:	心血管疾病;電學恆電位分析法;免疫電學感測;(導電型)原子力顯微術;cardiovascular disease;(conductive) atomic force microscopy;amperometry;label-free;electrical immunosensing;e- Ab sensor
日期:	2010
上传时间:	2010-09-23 16:12:16 (UTC+8)
摘要:	根據衛生署統計，97年國內十大主要死因(除意外事故外)中，心血管疾病造成死亡的人數占總死亡人數的11.1 %，僅次於惡性腫瘤的27.3 %。因此心血管疾病是當今不容忽視的慢性疾病。如何能夠提供精準度高的檢測數據，以提供醫師正確的診斷O目前科學家最棘手的課題。有別於過去的醫學檢驗報告，大部分都需透過實驗室大型儀器設備才能檢測得知，這種方式往往是曠日費時。因此，微型化檢驗機台之開發乃是定點照護最需要之設備。尤其現今國人高度注重健康以及社會已趨向高齡化，可攜式定點照護儀即變成不可或缺的配備。本研究針對主要心血管疾病標記分子，包含C-reactive protein (CRP)、cardiac troponin I (cTnI)以及S100A1 protein (S100A1)進行研究分析，並發展出一套新型無標定生醫免疫電學感測技術。此技術透過已設計並合成之有機電誘導型化合物分子進行電極晶片表面的自我組裝修飾；再將一系列心血管疾病標記分子進行生物固定化製程；配合電學感測技術之恆電位分析法於低電壓(≦-0.5V)下進行探討，研究發現抗體抗原間結合後形成一阻抗層並造成電極表面電性改變，藉由此變化可進行抗體抗原間專一性與非專一性之量測，且可量測達到ng等級之精確度。本研究並利用導電型原子力顯微技術觀察心血管疾病標記分子表面奈米結構與力學間之作用及電性變化。結果顯示，於低電壓(≦-0.5V)下，抗體抗原間之作用力不因電壓增加而改變，而高電壓(>-1V)下，有造成構形改變且抗體抗原間之作用力也隨之變化。再與X光結晶繞射法及螢光染色定量法之結果交叉比對，以提供此新型無標定生醫免疫電學感測技術更完整之技術架構。本技術之成功開發，有助於了解抗體抗原間之奈米結構、力學曲線與生物電性之關聯。並以此技術背景為主體，整合並研製完成微型化可攜式定點照護儀(e- Ab sensor)，結合微流道系統及可拋式免疫感測晶片，將即時顯示檢測結果，可提供醫師臨床診斷更完整且精準之檢驗數據，以縮短診斷時程並提升治癒效果。此裝置解決過去醫學檢驗使用實驗室大型儀器之困擾，提供未來居家自我看護更完善之微型化可攜式定點照護儀。未來，除了醫學檢驗可以更快速外，更進一步提供家庭、機場、醫院、車站等居家或公共場所的慢性疾病控制、用藥監測、環境稽查、食品檢驗以及毒品測試等等更完善的定點照護機制。 According to the statistics of Department of Health (DoH), the deaths caused by cardiovascular disease have 11.1 % in total number of deaths at Taiwan the year before last (2008). Currently, people have become more concerned about health and fitness. In the past, clinical diagnostic examination with blood required bulky equipments, time-consuming and expert staff in the hospital or central laboratory. Therefore, development of medical or healthcare monitoring device for detection and control of cardiovascular disease is in needed of point of care in the aging population. This work develops the novel electrical immunosensing technique for screening and analysis of major cardiovascular disease markers, including C-reactive protein (CRP), cardiac troponin I (cTnI), and S100A1 protein (S100A1). The technique consists of conductance-enhanced molecules for SAM formation, biological species immobilization and electrical immunosensing. Manipulation of the novel technique in amperometric analysis at the low voltage (≦-0.5V), the electricity or specificity were measured in antibody-antigen interactions and the accuracy achieve nanogram (ng) level. Furthermore, this work analyzed nano-mechanical structure, force and electrical distributions using conductive atomic force microscopy. The data compare with results of x-ray crystal diffraction method and enzyme-linked immunosorbent assay qualitative methods to complete the framework of this novel technique. This work integrates the novel electrical immunosensing technique to a handheld diagnostic device, called “e- Ab sensor”. The device consists of microfluidic system, immunoelectrodes and disposable cartridge. It has many benefits including rapid diagnosis, high accuracy, reliable, reproducible, affordable, robust and user-friendly, therefore, it can be performed on site. e- Ab sensor is a platform technique which can further develop for various applications, such as healthcare screening, chronic-diseases monitoring, environmental inspection, food-safety testing and bacteria or virus level determinations. e- Ab sensor could be used to quickly screen people at home, airport, hospital, and emergency clinic to control outbreaks of diseases such as SARS and H1N1 flu. However, the trends toward miniaturization and automation demand more advances in developing microfluidic system before e- Ab sensor can realize the commercial potential.
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