研究生: |
魏志安 Chih-an Wei |
---|---|
論文名稱: |
表面電漿於固定化葡萄糖酵素之量測 Surface Plasmon Biosensing for Immobilized GOD |
指導教授: |
徐世祥
Shih-Hsiang Hsu |
口試委員: |
林保宏
Pao-hung Lin 張勝良 Sheng-Lyang Jang 莊敏宏 Miin-Horng Juang |
學位類別: |
碩士 Master |
系所名稱: |
電資學院 - 電子工程系 Department of Electronic and Computer Engineering |
論文出版年: | 2014 |
畢業學年度: | 102 |
語文別: | 中文 |
論文頁數: | 61 |
中文關鍵詞: | 表面電漿共振 、葡萄糖氧化酵素 |
外文關鍵詞: | surface plasmon resonance, glucose oxidase |
相關次數: | 點閱:500 下載:0 |
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近年來,因為表面電漿共振(Surface plasmon resonance, SPR)技術具有高靈敏度特性,所以被廣泛的應用在生物感測技術,藥物的開發及篩選、食品檢測、環境汙染監測以及疾病的檢測等等。
在全世界,每小時就有一人死於糖尿病,對人類來說這是一種嚴重的疾病。血糖濃度的測量,是告訴我們一個人在有糖尿病前期或糖尿病的情況。正常人的血糖濃度為110毫克每分公升(毫克/分公升)或更低,糖尿病人濃度為126毫克/分公升或更高,糖尿病前期的人濃度為110毫克/分公升和125毫克/分公升之間。在這篇論文中,葡萄糖氧化酵素(glucose oxidase, GOD)固定化可有效提高血糖檢測的敏感性。
而在SPR量測技術中,最常見到的是角度調製(angle modulation)和波長調製(wavelength modulation),在此,我們以波長調製為主,因為與角度調制有關的步進馬達位置重複性不若現今光通訊波長來的精準,我們利用通訊波段 1520~1570 nm,因為對於波長調製而言,此波段較為靈敏。
在實驗開始前,我們先利用商業軟體 matlab 來模擬我們實驗所需的不同種類金屬與金屬厚度,以這為基礎下,我們選擇較為穩定的金,其中如果金在厚度 30 nm 、波長為 1550 nm 時為最佳,共振角在61.9度,反射系數為 0.0196。
實驗中,我們利用化學反應 MUA、EDC、NHS、PBS 等,將GOD固定在稜鏡的金屬表面,分別對於不同濃度葡萄糖做測量 10、100、200 mg/dL,其靈敏度為 0.0423 nm/(mg/dL),解析度為 0.023 mg/dL,與SPR光纖比較,光纖SPR解析度為 0.099 mg/dL,我們的SPR系統比SPR光纖系統的解析度小約4倍。
對於我們的SPR通訊波段而言,量測極限可達 0.01 mg/dL,與市面上販售的羅氏ACCU-CHEK 血糖機相比,我們的量測極限是羅氏ACCU-CHEK 血糖機的1000倍。
In recent years, the surface plasmon resonance (SPR) technology is demonstrated with high sensitivity and widely utilized in biosensing, drug development/screening, food testing, environmental pollution monitoring and disease detection.
One person passed away per hour from diabetes in the world, which is a serious disease for human beings. Blood glucose concentration measuring is the way to tell if one have pre-diabetes or diabetes. A normal glucose level is 110 milligrams per deciliter (mg/dL) or below, a diabetic level is 126 mg/dL or higher, and a pre-diabetes level is between 110 mg/dL and 125 mg/dL. In this thesis, the glucose oxidase (GOD) was utilized for immobilization for higher sensitive glucose sensing.
In the SPR characterization technique, two modulation approached are commonly used, angle and wavelength. Due to the angular resolution and repeatability from the step motor, the wavelength modulation was utilized for testing. The telecommunication wavelength range was then chosen because of its high sensitivity compared with visible light.
First of all, the commercial software, matlab, was used to simulate SPR phenomena with various metals and their thickness. In this thesis, the stable metal, gold, was simulated to illustrate the reflection coefficient of 0.0196 and resonance angle of 61.9 degree with 30-nm thickness at 1550-nm operating wavelength.
Glucose oxidase (GOD) on the prism gold metal surface was immobilized by chemical reaction, MUA, EDC/NHS, and PBS. The sensitivity and resolution from various glucose concentrations of 10, 100, and 200 mg/dL were showing 0.0423 nm/(mg/dL) and 0.023 mg/dL, respectively. Compared with the SPR glucose fiber sensor with the resolution of 0.099 mg/dL, our SPR was demonstrating at least 4 times better.
The sensing limit from our SPR sensor with telecommunication wavelength was showing 0.01 mg/dL with 1000 times better than Roche ACCU-CHEK blood glucose meter.
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