臺灣博碩士論文加值系統

本研究以γ-Fe2O3磁性奈米粒子為核心，利用迭代方法(Iteration method)將金奈米粒子沉積在γ-Fe2O3磁性奈米粒子表面上形成核殼結構(Core-Shell)之鐵核金殼磁性奈米粒子(γ-Fe2O3@Au NPs)，從X光繞射(XRD)、吸收光譜(UV-vis)、超導量子干涉儀(SQUID)、雷射粒徑分析儀(DLS)、穿透式電子顯微鏡(TEM)和能量色散X-射線光譜(EDS)觀察形貌後確定γ-Fe2O3磁性奈米粒子的表面上有包覆金層。之後我們將鐵核金殼磁性奈米粒子應用在阿茲海默症相關的Tau蛋白檢測，先把鐵核金殼磁性奈米粒子以官能基修飾，然後將Tau蛋白之抗體(Anti-Tau)鍵結在金殼上，得到具有檢測阿茲海默症的鐵核金殼磁性奈米粒子試劑，並且利用法拉第磁光系統檢測Tau蛋白不同濃度。由過去的成果已知，隨著抗原抗體的結合過程中磁性奈米粒子會團聚形成較大的磁簇，而造成法拉第旋轉角的改變，利用葡聚醣包覆磁性奈米粒子的檢測極限為10 ng/ml (10 ppb)。在此次的實驗中我們利用鐵核金殼磁性奈米粒子在特定波長(532 nm)下具有局部表面電漿共振(LSPR)特性與磁性奈米粒子相互作用，使法拉第旋轉角變化量增強，大幅提升檢測靈敏度，其偵測極限可達到1 pg/ml (1 ppt)。此方法除了簡單方便，並且能即時監控結合過程，是未來極具發展潛力的阿茲海默症早期檢測技術。

In this study, γ-Fe2O3 nanoparticles are used as the core and gold were deposited on the surface of γ-Fe2O3 nanoparticles by using the iteration method to form the γ-Fe2O3@Au nanoparticles. The X-ray diffraction (XRD), UV-visible spectrum (UV-vis), superconducting quantum interference device (SQUID), laser diffraction particle size analyzer (DLS), transmission electron microscopy (TEM) and energy dispersive spectrometers (EDS) were used to make sure the gold were actually coated on the γ-Fe2O3 nanoparticles. Then γ-Fe2O3@Au nanoparticles were applied to the detection of tau protein which is associated with Alzheimer's disease. First, the γ-Fe2O3@Au nanoparticles were modified by functional groups and then the Tau antibody (Anti-Tau) was bind on the surface of gold shell to obtain the detection reagents of Alzheimer's disease. The magneto-optical Faraday rotation system was used to detect different concentrations of Tau protein. According to previous work, when antibody conjugated with antigen, the magnetic nanoparticles were agglomerated to form larger magnetic clusters. So the angle of Faraday rotation would change during the process. The detection limit of the dextran coated magnetic nanoparticles was 10 ng/ml (10 ppb). In this experiment, we use γ-Fe2O3@Au nanoparticles on which the local surface plasmon resonance (LSPR) can be induced under specific 532 nm laser so that can interact with the magnetic nanoparticles to increase the Faraday rotation angle. The detection sensitivity is significantly improved, and the detection limit of tau protein can reach 1 pg/ml (1 ppt). This method is not only simple but also convient to monitor the combination process in real-time. Our research has a great development potential in early stage detection of Alzheimer’s disease.

致謝 i
摘要 ii
Abstract iii
目錄 iv
表目錄 vii
圖目錄 viii
第一章 緒論 1
1-1 前言 1
1-2 磁性奈米粒子 2
1-2-1 磁性物質介紹 2
1-2-2 磁性奈米材料合成 4
1-2-3 磁性奈米粒子於生醫檢測上之應用 5
1-3 金奈米粒子 7
1-3-1 金奈米粒子介紹 7
1-3-2 金奈米粒子於生物感測器上之應用 8
1-4 磁光效應 11
1-4-1 磁光效應簡介 11
1-4-2 表面電漿共振增強法拉第磁光效應 13
1-5 阿茲海默症(Alzheimer's disease) 17
1-5-1 阿茲海默症介紹 17
1-5-2 阿茲海默症病程發展 17
1-5-3 阿茲海默症診斷 18
1-5-4 Tau蛋白簡介 19
1-5-5 Tau蛋白檢測 20
1-6 研究目的 22
第二章 實驗原理及介紹 23
2-1 法拉第效應 23
2-2 表面電漿共振( Surface Plasmon Resonance，SPR ) 26
2-2-1 金屬平面上的表面電漿共振 26
2-2-2 金屬奈米粒子的局部表面電漿共振 26
第三章 實驗架構及量測方法 29
3-1 磁性奈米粒子 29
3-1-1 葡聚醣包覆磁性奈米粒子合成方法(Dextran-coated Fe3O4 NPs, DMP s) 29
3-1-2 純化DMPs (GFC, Gel filtration chromatography ) 30
3-1-3 Tau抗體修飾到葡聚醣包覆磁性奈米粒子 31
3-2 鐵核金殼磁性奈米粒子 32
3-2-1鐵核金殼磁性奈米粒子合成方法(γ-Fe2O3@Au NPs ) 32
3-2-2 Tau抗體修飾到鐵核金殼奈米粒子 33
3-3 特性分析 35
3-3-1 紫外/可見光分光光譜儀(UV-vis Spectrometers) 35
3-3-2 X光繞射儀(X-ray Diffraction, XRD) 36
3-3-3 奈米粒徑暨介面電位分析儀(Nanoparticle and Zeta Potential Analyzer) 37
3-3-4 穿透式電子顯微鏡(Transmission Electron Microscope, TEM) 38
3-3-5 超導量子干涉磁量儀(SQUID) 39
3-3-6 拉曼光譜(Raman Spectra) 40
3-4 系統量測及數據分析方法 41
第四章 實驗結果與討論 44
4-1葡聚醣包覆磁性奈米粒子之特性鑑定 44
4-1-1 X光繞射分析 44
4-1-2 粒徑及界面電位分析 44
4-1-3超導量子干涉磁量儀(SQUID) 46
4-2鐵合金殼磁性奈米粒子之特性鑑定 47
4-2-1 UV-vis光譜分析 47
4-2-2 X光繞射分析 48
4-2-3粒徑及界面電位分析 48
4-2-4超導量子干涉磁量儀(SQUID) 51
4-2-5穿透式電子顯微鏡 52
4-2-6 拉曼光譜 55
4-3 法拉第磁光系統量測 55
4-3-1 純Tau抗體與純Tau抗原測量 55
4-3-2 Tau抗體修飾葡聚醣包覆磁性奈米粒子磁光效應測量 56
4-3-3 Tau抗體修飾鐵核金殼磁性奈米粒子磁光效應測量 59
第五章 結論 65
第六章 參考文獻 66

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