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研究生:李怡青
研究生(外文):Lee yi ching
論文名稱:螢光粒子在生化分析上的初步應用
論文名稱(外文):Preliminary Study of Fluorescent Particles for Biochemical Analysis
指導教授:傅傳博
指導教授(外文):C. Bor Fuh
學位類別:碩士
校院名稱:國立暨南國際大學
系所名稱:應用化學系
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:104
中文關鍵詞:功能性奈米粒子
外文關鍵詞:functional nanoparticles
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本論文係關於製備功能性奈米粒子( functional nanoparticles )並結合免疫吸附分析法 ( immunosorbent assay )做生化分離上之應用研究。
本研究成功製備40nm、80 nm 及150nm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 nm、150 nm、200nm。
接下來,將鏈黴抗生素蛋白 (Streptavidin) 或免疫球蛋白 (IgG) 固定化在 SiO2 包覆四氧化三鐵磁性載體上,再將生物素 (d-Biotin) 或蛋白質A (Protein A) 固定在帶有螢光染料的載體上,結合微盤做簡單生化分析上之測試研究。初步探討磁性載體大小,及顆粒數目在生化分析上的可能應用。
此次研究主要是運用鏈黴抗生物素蛋白(Streptavidin)-生物素(Biotin)和免疫球蛋白G(IgG)-蛋白質A(Protein A)這兩對抗原與抗體之間在顆粒間的作用。實驗設計中,我們改變顆粒大小、及注入顆粒數的不同等條件來進行沉積試驗。在磁場的作用下,由於他們具有很強的專一性,帶有磁性顆粒的Streptavidin 和IgG 可以成功地分別與標示有Biotin 和Protein A 之螢光顆粒二氧化矽顆粒作用,可作進一步的分析。在控制組實驗的對照之下,充分表現出好的專一性與高度的選擇性。
The goal of this study is to synthesize functional magnetic particles and combine with immunosorbent assay for biochemical analysis.
The sizes of magnetic nanoparticles were 40nm, 80 nm and 150nm, and SiO2 coated Fe3O4 were 80nm, 150 nm and 200nm.
Streptavidin or IgG was bound onto Fe3O4@SiO2 particles. Biotin or protein A was covalently bound to fluorescent marker, and applied to magnetic separation in microplate. We studied experimental parameters of magnetic particles size、and particles number for immunoassay. Preliminary study of immunoassay between particles was studied to demonstrate the potential application of this method for biochemical analysis.
The interactions of streptavidin-biotin and IgG-protein A that coating on particles was studied. Experimental parameters were studied under different conditions of the particle size and the number of particles. The streptavidin-conjugated magnetic
beads and IgG-conjugated magnetic beads can capture fluorescent silica particles labeled with biotin and protein A in the 96 well plate under magnetic filed. The selectivity is high with controlled experiments.
中 文 摘 要 I
目 錄 III
表 目 錄 VI
圖 目 錄 VII
第 一 章 緒 論 1
1.1 研究動機 1
1.2 二氧化矽 3
1.2.1二氧化矽的應用 3
1.2.2膠體之穩定機構與二氧化矽奈米粒子生成機制 4
1.3 磁性材料 11
1.3.1磁性奈米粒子 12
1.4 奈米技術在螢光檢測上的應用 14
第 二 章 功能性奈米粒子之製備 17
2.1 前言 17
2.2 實驗藥品與器材 19
2.3 實驗步驟 21
2.3.1 磁性奈米粒子之製備 21
2.3.2 二氧化矽奈米粒子之製備 21
2.3.3 磁性奈米粒子包覆二氧化矽之製備 21
2.3.4 二氧化矽螢光奈米粒子之製備 21
2.3.5 修飾磁性奈米粒子之製備 23
2.3.6 修飾螢光奈米粒子之製備 23
2.3.7 自製磁性顆粒送測後之計算處理 24
2.4 實驗結果 25
2.4.1 奈米粒子的大小及結構 25
2.4.2 修飾奈米粒子表面 35
2.4.3 磁性 40
2.4.4 表面結構及化學組成 46
2.4.5 Rhodamine 6G包覆二氧化矽螢光強度 48
2.4.6 Rhodamine 6G包覆二氧化矽螢光穩定性 50
第 三 章 牛血清蛋白 (BSA) 及抗牛血清蛋白 (anti-BSA) 在分析磁盤技
術上的作用 51
3.1 前言 51
3.2 實驗藥品與器材 56
3.3 實驗步驟 57
3.4 結果與討論 63
第 四 章 鏈黴抗生物素蛋白(Streptavidin)及生物素(Biotin)在分析磁
盤技術上的作用 68
4.1 前言 68
4.2 實驗藥品與器材 70
4.3 實驗步驟 71
4.4 結果與討論 77
第 五 章 蛋白質A(Protein A)及免疫球蛋白G(IgG)在分析磁盤技術
上的作用 83
5.1 前言 83
5.2 實驗藥品與器材 88
5.3 實驗步驟 89
5.4 結果與討論 94
第 六 章 結論與未來展望 99
6.1 結論 99
6.2 未來與展望 100
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