阿茲海默症是一種會導致記憶力衰退和其他認知缺陷的神經退化性疾病，在六十五歲以上的老年人中，約有一成的人患有阿茲海默症，其形成的主要原因與β類澱粉蛋白寡聚物的堆疊有關。因此，β類澱粉蛋白在阿茲海默症的研究中扮演重要的一環。在本論文中，我們開發了一種新型的螢光標記探針，可在複雜的環境中具有選擇性的偵測β類澱粉蛋白，此探針展現了高靈敏度及高穩定性。在此探針中，我們將硫代硫黃素的衍生物作為結合配體，並引入二氟苯酚酯作為反應端，當配體與β類澱粉蛋白產生專一性親和力結合後，目標蛋白表面的親核基會與探針反應並形成共價鍵，可有效標記β類澱粉蛋白。我們相信此探針將在β類澱粉蛋白和阿茲海默症的研究中扮演非常有用的工具。

Alzheimer’s disease (AD) is a neurodegenerative illness, causing memory loss and other cognitive deficits. It affects about 10% of older adults aged 65 years and older. The main cause of AD is widely believed to be associated with the formation of β-amyloid protein oligomers. Therefore, the study of β-amyloid protein is an important topic in AD research. In this thesis, we introduce a novel fluorescent labeling probe for the selective modification of β-amyloid in a complex biological environment. The probe exhibits excellent sensitivity and high stability. The probe was created by the incorporation of a Thioflavin T derivative as the binding ligand and difluorophenyl ester as the reactive group to facilitate efficient labeling on β-amyloid. After the specific binding to β-amyloid, the nucleophilic residues of amino acid on β-amyloid can react with the ester group to form a covalent bond to effectively label the target. We believe that the probe would be a very useful tool for the study of β-amyloid and Alzheimer’s disease.

摘要 I
Abstract II
謝誌 III
目錄 V
第一章 緒論 1
1-1蛋白質 1
1-2 類澱粉蛋白 2
1-3 阿茲海默症 3
1-4蛋白質偵測方法 5
1-4-1 高效能液相層析法 5
1-4-2 質譜分析法 6
1-4-3 西方墨點法 7
1-4-4 酵素結合免疫吸附分析法 8
第二章 文獻回顧 11
2-1 𝛽類澱粉蛋白的體外偵測 11
2-1-1 電化學技術 11
2-1-2 懸臂式脂質生物感測器技術 13
2-1-3 酵素結合免疫吸附分析法技術 14
2-1-4 質譜技術 15
2-2 𝛽類澱粉蛋白的體內偵測 16
2-2-1 正電子斷層掃描 16
2-2-2 奈米粒子與電化學技術 18
2-2-3 微透析技術 19
2-2-4 碳纖維微電極技術 20
2-3 螢光探針偵測𝛽類澱粉蛋白 21
2-3-1 硫黃素T衍生物螢光探針 21
2-3-2 薑黃素衍生物螢光探針 22
2-3-3 BODIPY衍生物螢光探針 23
第三章 探針設計 24
3-1 螢光探針之構想 24
3-2 硫黃素 25
3-3 螢光探針之設計 26
第四章 實驗結果與討論 27
4-1探針1 27
4-1-1 探針1之合成 27
4-1-2 探針1實驗結果 28
4-2探針3 29
4-2-1 探針3之合成 30
4-2-2 探針3與β-amyloid的標記實驗 32
4-3探針4 34
4-3-1 探針4之合成 34
4-3-2 探針4與β-amyloid的標記實驗 37
4-4探針2 39
4-4-1 探針2之合成 40
4-4-2 探針2實驗結果 41
第五章 實驗實驗結論 45
第六章 實驗部分 46
6-1一般驗藥品與器材 46
6-2 十二烷基硫酸鈉聚丙烯醯胺凝膠電泳 48
6-2-1 SDS-PAGE膠體電泳蛋白質標準液 48
6-2-2 SDS-PAGE膠體電泳配方 48
6-2-3 蛋白質的反應流程 49
6-2-4 SDS-PAGE膠體電泳流程 50
6-2-5 銀染色法流程 50
6-2-6膠體螢光強的定量 51
6-3 有機合成及光譜資料 52
第七章 參考資料 84
附錄 90

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