臺灣博碩士論文加值系統

本論文係關於製備功能性磁性粒子( functional magnetic particles )並結合分析磁泳
技術( analytical magnetapheresis )做生化分離上之應用研究。前者主要利用化學共沉
澱方法( co-precipitating )製備磁性粒子，再對磁性粒子以高分子修飾粒子表面，所修
飾磁性粒子表面之官能基有NH2、COOH，最後採用穿透式電子顯微鏡( transmission
electron microscope；TEM )、掃描式電子顯微鏡( scanning electron microscope；SEM )、
粒徑分析儀( photon correlation spectroscopy；PCS )、原子力顯微鏡( atomic force
microscope；AFM )、超導量干涉磁量儀( superconducting quantum interference device
(SQUID) magnetometor)、X射線繞射儀( x-ray diffractometer；XRD )、傅立葉轉換紅
外線光譜儀( Fourier transform infrared instrumentents；FT-IR )、紫外光/可見光
( UV-VIS )鑑定。本研究成功製備40 ± 3 nm、100 ± 8 nm 及300 ± 14 nm之Fe3O4粒子。
磁性粒子表面經有機物質修飾粒子表面後粒徑為80 ± 5 nm、200 ± 15 nm、420 ± 38
nm、860 ± 52 nm及2000 ± 112 nm。
接下來，將鏈黴抗生素蛋白(Streptavidin)或免疫球蛋白(IgG)固定化在四氧化三鐵/
高分子磁性載體上，再將生物素(d-Biotin)或蛋白質A(Protein A)固定在二氧化矽( SiO2 )
載體上，結合分析磁泳技術( analytical magnetapheresis )做簡單生化分離上之測試研
究。探討磁性載體種類、流速、及顆粒數目在分離上的可能應用。

Analytical magnetapheresis has become a useful technique for analyzing magnetically
susceptible particles. Magnetic particles ( Fe3O4 ) were prepared by co-precipitating.
The surface modifications of Fe3O4 were studied with polymer and NH2, COOH.
Transmission electron microscopy ( TEM ) , photon correlation spectroscopy ( PCS ) ,
atomic force microscope ( AFM ) , x-ray diffraction ( XRD ) , superconducting quantum
interference device (SQUID) magnetometer , fourier transform infrared instruments
( FT-IR ) and UV-VIS were used for analysis. The sizes of magnetic nanoparticles were :
Fe3O4 = 40 ± 3 nm、100 ± 8 nm and 300 ± 14 nm , and Fe3O4-polymer= 80 ± 5 nm、200 ± 15
nm、420 ± 38 nm、860 ± 52 nm and 2000 ± 112 nm。
Streptavidin or IgG was bound onto Fe3O4-polymer particles via carbodiimide
activation. Biotin or IgG was covalently bound SiO2 particles, and applied to magnetic
carriers for bioseparation in analytical magnetapheresis. We studied experimental
parameters of magnetic particles、flowrate and particles number for analytical
magnetapheresis.

中文摘要.................................................................................................................... I
英文摘要................................................................................................................... II
目錄……………………………………………………………………………………...III
表目錄…………………………………………………………………………………VI
圖目錄…………………………………………………………………………….........IX
第一章緒論...........................................................................................................1
1.1 前言.................................................................................................................... 1
1.2 研究動機............................................................................................................ 3
1.3 奈米材料分類與製備......................................................................................... 4
1.4 奈米材料表面修飾............................................................................................. 7
1.5 功能性磁性奈米材料....................................................................................... 10
1.5.1 氧化鐵之特性........................................................................................ 10
1.5.2 磁性材料應用........................................................................................ 11
1.6 磁性分析技術之原理....................................................................................... 14
第二章理論.........................................................................................................16
2.1 磁性相關理論.................................................................................................. 16
2.1.1 磁化率之觀念與換算............................................................................ 20
2.1.2 磁化率之量測........................................................................................ 23
2.1.3 超導量子干涉磁量儀測量法( SQUID magnetometer ) ......................... 23
2.2 高分子分散聚合的發展與原理....................................................................... 29
2.2.1 高分子合成........................................................................................... 29
2.2.2 分散聚合法........................................................................................... 23
2.3免疫分析之原理............................................................................................... 34
2.3.1 非專一性免疫........................................................................................ 34
2.3.2 專一性免疫反應.................................................................................... 36
2.3.3 抗體與抗原之結合力............................................................................ 37
2.4分析磁泳技術之原理........................................................................................ 40
2.4.1磁場( magnetic field ) ............................................................................. 40
2.4.2分離槽( separation channel )................................................................... 47
2.4.3多管注射幫浦( syringe pump ) ............................................................... 47
第三章功能性磁性粒子之製備............................................................................48
3.1 前言.................................................................................................................. 48
3.2 實驗藥品與器材.............................................................................................. 50
3.2.1 實驗藥品............................................................................................... 50
3.2.2 儀器....................................................................................................... 51
3.2.3 材料....................................................................................................... 51
3.2.4 自製磁性顆粒送測後之計算處理......................................................... 51
3.3 實驗步驟.......................................................................................................... 53
3.3.1 磁性奈米粒子之製備............................................................................ 53
3.3.2 無孔性高分子顆粒製備........................................................................ 53
3.3.3 修飾磁性奈米粒子之製備.................................................................... 53
3.4 特性分析.......................................................................................................... 57
3.4.1 穿透式電子顯微鏡( TEM )分析............................................................ 57
3.4.2 掃描式電子顯微鏡( SEM )分析............................................................ 57
3.4.3 原子力顯微鏡( AFM )分析................................................................... 57
3.4.4 雷射粒徑儀分析.................................................................................... 57
3.4.5 X射線繞射儀( XRD )分析.................................................................... 57
3.4.6 傅立葉轉換紅外線光譜儀( FTIR )分析................................................ 57
3.4.7 紫外光/可見光光譜儀( UV-VIS )分析.................................................. 57
3.5 實驗結果.......................................................................................................... 58
3.5.1 磁性奈米粒子大小及結構.................................................................... 58
3.5.2 修飾磁性奈米粒子表面........................................................................ 61
3.5.3 磁性....................................................................................................... 64
3.5.4 磁性次微米粒子大小及結構................................................................ 69
3.5.5 粒子對紫外光/可見光( UV-VIS )吸收探討.......................................... 74
第四章鏈黴抗生素蛋白(Streptavidin) 及生物素(d-Biotin)在磁性分離技術上
的應用.......................................................................................................76
4.1 前言.................................................................................................................. 76
4.2 實驗藥品與器材.............................................................................................. 78
4.2.1 實驗藥品............................................................................................... 78
4.2.2 儀器....................................................................................................... 78
4.3 實驗步驟.......................................................................................................... 79
4.3.1 磷酸鹽緩衝溶液(PBS)之配製............................................................... 79
4.3.2 血球計數盤使用方法............................................................................ 80
4.3.3 樣品配製............................................................................................... 81
4.4 實驗結果.......................................................................................................... 86
4.4.1 不同磁性顆粒大小對鍵結鏈黴抗生素蛋白(Streptavidin)之影響................ 86
4.4.2 流速之影響................................................................................................... 86
4.4.3注入不同數目二氧化矽鍵結生物素(d-Biotin)顆粒數的影響....................... 86
4.4.4磁珠固定鏈黴抗生素蛋白(Streptavidin)添加NHS之影響........................... 94
第五章免疫球蛋白(IgG)及蛋白質A(Protein A)在磁性分離技術上的應用....... 102
5.1 前言................................................................................................................ 102
5.2 實驗藥品與器材............................................................................................ 103
5.2.1 實驗藥品............................................................................................. 103
5.2.2 儀器..................................................................................................... 103
5.3 實驗步驟........................................................................................................ 104
5.3.1磷酸鹽緩衝溶液(PBS)之配製.............................................................. 104
5.3.2 樣品配製............................................................................................. 105
5.4 實驗結果.........................................................................................................110
5.4.1不同磁性顆粒大小對鍵結免疫球蛋白(anti-rabbit IgG)之影響....................110
5.4.2 流速之影響..................................................................................................110
5.4.3 顆粒數之影響..............................................................................................110
5.4.4磁珠固定免疫球蛋白(anti-rabbit IgG)添加NHS之影響.............................119
第六章結論與未來展望...................................................................................... 127
參考文獻................................................................................................................ 129

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