臺灣博碩士論文加值系統

近年來，由於晶片技術的成熟發展，生醫感測晶片應用於臨床醫學檢測疾病上的地位也越來越重要。然而在製作生醫感測晶片過程中，利用自我組裝單分子層來固定蛋白質已經是目前蛋白質固定技術的主流之一。儘管不斷的努力，自我組裝單分子層（self assembled monolayer, SAM）緻密程度與金電極表面的潔淨度、均勻度皆會導致蛋白質無法均勻固定於金電極上，使得生醫感測晶片的靈敏度大幅下降，因此本研究以11-mercaptoundecanoic acid (MUA)與金電極表面進行自我組裝單分子層之特性評估，並以電化學阻抗分析法(electrochemical impedance spectroscopy, EIS)和循環伏安法(cyclic voltammetry, CV)、親疏水角量測儀(contact angle goniometry)進行分析，研究結果顯示最佳修飾條件在奈米晶片濺鍍金以功率80W 電流 20mA 濺鍍時間120 秒可在奈米聚碳酸脂(Polycarbonate)產生均勻的奈米金顆粒薄膜；自我組裝單分子層利用浸泡法(immerse method)並利用電化學阻抗分析法找出MUA濃度500M 3小時即可達到緻密的自我組裝單分子層，並使生醫感測晶片遮蔽層(Blocking)阻抗重複性之變異值<10%，並應用於過敏性物質組織胺(Histamine)量測濃度範圍1ng/ml ~ 1000ng/ml之線性度可達 0.9851，與Tryptase量測濃度範圍0.1ng/mL ~ 50 ng/mL之線性度可達 0.9828。

Despite considerable efforts, the fabrication of protein chips using a self-assembled
monolayer (SAM) with a long chain length remains a challenge, due to its substrate cleanliness and evenness. Therefore, the protein can not be stably fixed on the substrate, resulting in a significant drop in the sensitivity of the bio-sensors. This study improved the repeatability of self-assembled monolayer on chip. We use self-assembled monolayer (SAM) with functional groups to modify a substrate. Thus the surface of the substrate is suitable for specific bio-detection. The two characteristics of this study, one characterize is that the physical properties of substrate with SAM modification, the other is that applied a layer of SAM coated with protein on sensing electrode as a bio-probe to quantify the formation of avidin-biotin complex by impedance response in electrical impedance analysis.It was less than 10% of the repeatability of resistances 〖 R〗_bsa in Histamine and Tryptase, after Protein incubate on sensors. ΔR is the definition of R_protein minus〖 R〗_bsa. The Histamine is measured by EIS concentration measurement which resulted a linear equation of Y=4107ln(X) +5681.6 with error R^2 = 0.985 in impedance change. The Tryptase is measured by EIS concentration measurement which resulted a linear equation of Y=702.91ln(X) +17332 with error R^2 = 0.9828 in impedance change.

誌謝 i
摘要 ii
ABSTRACT iii
目錄 iv
圖目錄 vi
表目錄 viii
第一章、 前言 1
1.1. 研究動機 1
1.2. 研究目標 9
1.3. 論文架構 9
第二章、 相關理論與技術 10
2. 1 電化學 10
2.1.1 電化學質傳與電子交換現象 10
2.1.2 電極程序之動力學 12
2.1.3 界面現象與電極雙層模型 13
2. 2循環伏安法 15
2. 3 電化學阻抗分析法 18
2.3.1 電化學阻抗分析等效電路 18
2.3.2電化學阻抗分析動態化學過程 19
2.3.3 電化學阻抗分析阻抗參數 21
2. 4抗體抗原免疫分析 23
2.4.1抗體 23
2. 5微奈米轉印技術 24
2. 6表面分析技術 24
2.6.1接觸角(Contact Angle) 24
2.6.2掃描式電子顯微鏡(Scanning Electron Microscope，縮寫為SEM) 25
2.6.3原子力顯微鏡(Atomic force microscope，AFM) 25
第三章、 實驗材料與方法 26
3. 1 調配自我組裝單分子層溶液 26
3. 2奈米陣列檢測晶片 26
3.2.1 3D奈米結構 28
3.2.2米3D半球結構AAO模型製作 30
3.2.3熱壓奈米半球PC試片 30
3.2.4鍍金電極 32
3.2.5奈米陣列檢測晶片封裝 33
3.3 自我組裝單分子層(self-assembly monolayers) 34
3.4 電化學阻抗量測 36
3.5 實驗藥品、設備 37
第四章、 結果與討論 38
4.1 奈米半球結構AAO模具 38
4.2 奈米半球結構PC試片 39
4.3 奈米半球結構PC鍍金 40
4.4 電化學治具 41
4.5 金電極確認 43
4.6 自我組裝單分子層 44
4.6.1 濃度對自我組裝薄影響 44
4.6.2 時間對自我組裝單分子層影響 47
4.6.3 MUA drop method V.S. immerse method 47
4.6.4 MUA自我組裝單分子層接觸角 49
4.6.5 藉由EDC/NHS活化自我組裝單分子層 50
4.7 等效電路模型建立 53
4.8 自我組裝單分子層應用於Histamine 阻抗分析 53
4.8.1檢測晶片抗體最佳接附量 54
4.8.2 Histamine標準曲線 56
4.9 自我組裝單分子層應用於Tryptase 阻抗分析 59
4.9.1 Tryptase標準曲線 59
第五章、 結論與未來展望 63
5.1 結論 63
5.2 未來展望 64
參考文獻 65

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