臺灣博碩士論文加值系統

細胞在進行呼吸或免疫反應時，會產生活性氧物質（reactive oxygen species, ROS）；當體內過量ROS生成累積時，會形成氧化壓力（oxidative stress），造成心血管與神經病變，如帕金森症、類風濕性關節炎等，因此開發可即時檢測ROS之生物感測器為當今研究趨勢。本研究電沉積不同奈米金結構（gold nanostructure, GNS）於金薄膜電極上，並直接吸附或透過3-mercaptoproponic acid (MPA)修飾後再吸附超氧化物歧化酶（superoxide dismutase, SOD）於GNS上，以構成超氧陰離子(O_2^(∙-))感測電極。實驗中探討表面零電荷電位（potential of zero charge, pzc）、GNS晶格面與形狀對SOD的覆蓋率、電子轉移的影響。實驗結果證實，直接型吸附SOD電極的陰極轉移係數(α_c)與電子轉移速率(ks)受晶格面影響，其中(200)面有助於α_c與ks之反應，而SOD覆蓋率隨粗糙度增加而增加。當以MPA當促進劑（promoter）以靜電力吸附SOD時，在相同GNS電極上，pzc越正，越能促使SOD陰極反應的發生，而SOD覆蓋量則與MPA吸附量呈正相關。
以電化學參數最佳之SOD/ MPA(10 mM修飾24 h)/GNS(1+2+3)電極（SOD覆蓋量為8.79×〖10〗^(-11) mol cm^(-2)，α_c與ks分別為0.66與254.1 s-1），對超氧陰離子進行檢測時，其靈敏度與線性區間分別為1.99 nA μM^(-1) cm^(-2)與0.52-1.73 μM，本研究已顯示成功開發一可量測O_2^(∙-)之第三代SOD電極。

When cells perform the behaviors of respiration or immunization, reactive oxygen species (ROS) are produced. An excessive accumulation of ROS forms the oxidative stress for an organism, causing Parkinson’s, cardiovascular disease and rheumatoid arthritis. Therefore, it is one of the tendencies to develop a biosensor to reach the real-time detection of SOD. In this study, different gold nano-structures (GNS) were electrodeposited on the thin-film gold electrodes. Subsequently, superoxide dismutase (SOD) was directly adsorbed on GNS or on 3-mercaptoproponic acid (MPA)-modified GNS to construct a superoxide anion (O_2^(∙-))-sensing electrode. The effects of the potential of zero charge (pzc), the GNS facets and morphology on the SOD coverage and the electron transfer properties are discussed. The results show that cathodic transfer coefficient (αc) and electron transfer rate (ks) were affected by the GNS facets at the SOD direct-absorption electrodes. The (200) facet could facilitate the reaction of αc and ks, and the SOD coverage increased with the surface roughness. In terms of MPA-promoter SOD electrodes, the more positive pzc promoted more cathodic reaction of SOD. Moreover, the SOD coverage was proportional to the MPA coverage.
The SOD/MPA(10 mM for 24 h modification)/GNS(1+2+3) electrodes presented the highest SOD coverage of about(8.79 ± 0.63)×〖10〗^(-10) mol cm^(-2) and the largest αc of 0.66 and ks of 254.1 s-1. The sensitivity and linear range was 1.99 nA μM^(-1) cm^(-2) and 0.52–1.73 μM. The study proves that a third-generation SOD electrode has been successfully achieved for O_2^(∙-) detection.

摘要 I
Abstract II
圖目錄 VII
第一章 緒論 1
1.1研究背景 1
1.2細胞型生物感測器的定義、結構與發展 3
1.2.1細胞型生物感測器的定義 3
1.2.2細胞活性評估之生理意義 4
1.2.3 活性氧物質於體內之來源、影響及檢測技術 6
1.3超氧陰離子酵素感測器 13
1.3.1超氧陰離子安培法酵素感測器原理 13
1.3.2 超氧陰離子之細胞氧化壓力評估 17
1.4電化學製備金奈米結構的方法 18
1.4.1 底材效應 18
1.4.2 保護劑的運用 30
1.4.3 晶種成長（seed-mediated growth） 35
1.5 結合GNS之超氧陰離子酵素感測器 37
1.6表面電位對酵素的影響 40
1.7 研究目的與架構 42
第二章 實驗理論分析 44
2.1電極表面零電荷電位量測原理 44
2.2 酵素反應與酵素電極 50
2.2.1酵素動力學 50
2.2.2 酵素固定技術 52
第三章 材料與方法 54
3.1實驗設備 54
3.2 實驗藥劑 56
3.3感測晶片設計 60
3.4感測晶片製作 61
3.4.1 玻璃清潔 61
3.4.2 正光阻Lift-off製作 61
3.4.3絕緣層製作 63
3.4.4電沉積GNS結構 64
第4章 結果與討論 66
4.1 晶片架構 66
4.2 GNS電化學特性與表面結構 67
4.2.1 GNS結構特性 67
4.2.2 GNS電極電化學特性 75
4.2.3 奈米金表面結構特性 80
4.2.4 Potential of zero charge 分析GNS表面電位 82
4.3 SOD直接吸附型電極之電化學特性 84
4.3.1 SOD/GNS酵素電極電化學特性 85
4.3.2 SOD吸附於經MPA修飾之酵素電極 90
4.3.3 SOD-MPA-GNS酵素電極電化學反應特性整理 100
4.3.4 SOD酵素電極耐久性（durability） 101
4.4 SOD酵素電極O_2^(∙-)量測反應 105
第5章 結論 112
參考文獻 113

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