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系統識別號 U0026-1507201610035600
論文名稱(中文) 以交流電驅動促進之電化學免疫晶片偵測心肌肌鈣蛋白 I 的研發
論文名稱(英文) Study on AC Electro-kinetics Enhanced Electrochemical Immuno-chip to Detect Cardiac Troponin I
校院名稱 成功大學
系所名稱(中) 生物醫學工程學系
系所名稱(英) Department of BioMedical Engineering
學年度 104
學期 2
出版年 105
研究生(中文) 程玉
研究生(英文) Yu Cheng
學號 S46034035
學位類別 碩士
語文別 英文
論文頁數 60頁
口試委員 指導教授-張憲彰
口試委員-莊漢聲
口試委員-丁信智
口試委員-陳之碩
口試委員-陳立家
中文關鍵字 心肌肌鈣蛋白 I  交流電滲流  方波伏安法  電化學阻抗分析法 
英文關鍵字 Cardiac Troponin I (cTnI)  Alternating current electro-osmosis (ACEO)  Square wave voltammetry (SWV)  Electrochemical impedance spectroscopy (EIS) 
學科別分類
中文摘要 冠狀動脈疾病為造成全球致死的主因之一,隨著罹患病患數量大幅增加,檢測的方便性及快速性是亟需克服的重要課題。近期廣泛使用的免疫檢測法,雖可以達到靈敏及準確的目標,但是傳統的免疫檢測法需要使用螢光標的及酵素呈色反應,與抗原利用擴散作用和抗體結合,而使檢測時間冗長,最後需要配合大型光學儀器分析。因此,快速、靈敏、低成本及簡便的檢驗方法便為生醫檢測器的目標。
為了增加靈敏度及偵測極限,結合電動力學交流電滲流 (ACEO) 及不須標記之電化學偵測可藉流體對流以增加抗體-抗原反應。此外,本研究藉最佳化金電極之修飾實驗流程以建立良好再現性,例如:自主性單層膜、化學鏈結劑、抗體及阻滯劑。多種電化學偵測法包含循環伏安法、方波伏安法及電化學阻抗分析法被用於定量心肌梗塞指示劑-心肌肌鈣蛋白蛋白I (cTnI)。
實驗結果顯示,在同心圓電極施加偏壓0.5 Vpp、內圈電壓8 Vpp及外圈電壓15 Vpp,與最佳頻率800 Hz的條件下,能產生最佳濃縮效果,不僅能減少反應時間至20秒,且藉由短暫的暫停造成的再次攪拌能增加代測物的結合率,以期未來能結合自動化,達到更快速檢測之效果。
英文摘要 Myocardial infarction (MI) is mostly induced by coronary artery disease (CAD) which is one of the major lethality worldwide and it is predicted to rise. Although conventional immunoassays achieve good sensitivity and specificity, the long reaction time and bulky optical instruments are needed. Therefore, the sensing performances involving fast, high sensitivity, low cost and reproducibility are critical for the development of biosensors.
For increasing the sensitivity and decreasing the reaction time, the integration of electrokinetic alternating current electro-osmosis (ACEO) into label-free electrochemical detection could facilitate antibody-antigen reaction by driving fluidic convection. Furthermore, a good reproducibility was achieved by optimizing the procedures of surface modification on gold electrode, such as self-assembled monolayer, chemical linker, antibody, and blocker. Various electrochemical detections consisting of cyclic voltammetry (CV), square wave voltammetry (SWV), and electrochemical impedance spectroscopy (EIS) were used to quantify cardiac troponin I (cTnI) which is an important indicator of MI.
The data showed that applying concentric electrode optimal frequency 800 Hz, inner 8 Vpp and outer 15 Vpp with bias 0.5 Vpp to generate the concentrated effect which can eliminate reaction time from 15 min to 20 s, and with a pause enhances the binding rate of analyte due to the re-stirring. With the prospect of automation, faster and more accurate detection can be achieved.
論文目次 Abstract I
摘要 II
致謝 III
Contents IV
List of Figures VI
List of Tables XI
Chapter 1 Introduction 1
1.1 Background and Motivation 1
1.2 Biosensors 4
1.2.1 Conventional Immunosensors 5
1.2.2 Electrochemical Biosensors 6
1.3 Micro-Electro-Mechanical System Technology 7
1.4 Theory of Electrokinetic 8
1.4.1 Electric Double Layer 8
1.4.2 Dielectrophoresis (DEP) 11
1.4.3 Electro-osmosis (EO) 12
1.5 Electrochemical Detection Methods 17
1.5.1 Cyclic Voltammetry (CV) 17
1.5.2 Square Wave Voltammetry (SWV) 18
1.5.3 Electrochemical Impedance Spectroscopy (EIS) 19
1.6 Research Framework 24
Chapter 2 Materials and Methods 26
2.1 Microchip Design and Fabrication 26
2.2 Apparatus 28
2.3 Reagents and Solutions 28
2.4 Modification and Detection Procedure 29
2.5 Electrochemical Measurements 31
2.6 Statistic Formula 31
2.7 System Configuration 31
Chapter 3 Results and Discussion 33
3.1 Verification for Conditions of Surface Modification 33
3.1.1 Reproducibility of Commercial Electrodes 33
3.1.2 Immobilization of 11-MUA 35
3.1.3 Modification of EDC/NHS 40
3.1.4 Immobilization of the Anti-cTnI Antibody 42
3.1.5 Sensing Performances for cTnI Detection 45
3.2 Verification for Applying ACEO Conditions 48
3.2.1 Reproducibility of ACEO Electrodes 49
3.2.2 Evaluation for Applying Time of ACEO 50
3.2.3 Demonstration of Sensitivity Improvement under ACEO 52
3.2.4 Effects of ACEO Application Duration 52
Chapter 4 Conclusion and Prospect 54
Acknowledgement 55
References 56
Curriculum Vitae 60
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