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論文中文名稱:以奈米金及免疫微米磁珠檢測半抗原之研究 [以論文名稱查詢館藏系統]
論文英文名稱:Hapten detection using gold nanoparticles and immunomagnetic microbeads [以論文名稱查詢館藏系統]
院校名稱:臺北科技大學
學院名稱:工程學院
系所名稱:化學工程與生物科技系生化與生醫工程碩士班
畢業學年度:105
畢業學期:第一學期
出版年度:106
中文姓名:陳宜鄉
英文姓名:Yi-Shiang Chen
研究生學號:102688020
學位類別:碩士
語文別:中文
口試日期:2014/07/09
論文頁數:43
指導教授中文名:侯劭毅
指導教授英文名:Shao-Yi Hou
口試委員中文名:黃志宏;劉正哲;王勝仕
口試委員英文名:Chih-Hung Huang;Cheng-Che Liu;Sheng-Shi Wang
中文關鍵詞:免疫磁珠奈米金粒子磁性分離法半抗原
英文關鍵詞:immunomagnetic microbeadsgold nanoparticlesimmunomagnetic separation analysishapten
論文中文摘要:本實驗將奈米金標記與免疫磁珠結合,藉磁性分離檢測半抗原。原理為利用微米磁珠上修飾抗生物素抗體,以膠體金為標記物鍵結定量抗原,抗體與奈米金上之抗原產生專一性結合反應,再透過強力磁鐵吸附磁珠,將結合磁珠上之膠體金與未結合之膠體金分離,偵測殘留膠體金其吸收峰值取得校正曲線。在磁珠上修飾多株抗體,因免疫反應捕獲膠體金上修飾之目標物,造成反應液中膠體金數量減少,因而導致顏色由紅色漸減至無色;當樣品含有目標物,則會與膠體金上目標物競爭抗體接合位,殘留更多膠體金使溶液呈現紅色。使用兩種不同之奈米金探針,Biotin-DNA奈米金探針在1 pmol至10nmol有兩個良好線性區間,檢出偵測極限為1 pmol,Au-biotin奈米金探針則在5 pmol至50 nmol,偵測極限為5 pmol,操作方便並且可於20分鐘檢測完成。
論文英文摘要:In this study, we used nanogold labeling and immunomagnetic beads with magnetic separation to detect hapten. Micro magnetic beads modified antibodies which capture target immobilized on colloidal gold. To separate colloidal gold conjugated to magnetic beads from solution by magnet and uncombined AuNPs could be directly quantified by UV–Vis absorption spectroscopy of the supernatant. Polyclonal antibodies modified on the beads would have immune response with targets , causing the reaction solution colors fading from red to colorless. If the sample had the targets , which would compete the binding sites of antibodies with targets immobilized on colloidal gold. Using two kinds of nanogold probe detection biotin . One is biotin-DNA nanogold probe and the calibration curve contains two linear ranges. The equations for the linear fits between 1pmol and 100pmol and between 100pmol and 10nmol. The limit of quantitation (LOQ) value was 1 pmol. Other one is Au-biotin nanogold probe and the calibration curve contains two linear ranges, too. The LOQ value was 5pnol. This detection system is operated easily and it takes 20 min to obtain the results.
論文目次:摘要 I
ABSTRACT II
目錄 V
圖目錄 VII
第一章 緒論 1
1.1 前言 1
1.2 研究目的 1
第二章 研究背景與實驗原理 2
2.1 半抗原(Hapten)簡介 2
2.2 現行標準方法檢測半抗原之介紹 3
2.3 競爭法檢測半抗原之原理介紹 6
2.3.1 競爭抑制酵素免疫分析法 6
2.3.2螢光偏振免疫分析法 8
2.3.3奈米金增強酵素免疫法 9
2.3.4 比色法 10
2.3.5免疫層析法 14
2.3.6表面電漿共振生物感測器 17
2.2 本實驗之發展奈米金探針-以奈米金及免疫磁珠檢測半抗原 21
第三章 實驗方法與步驟 22
3.1實驗架構 22
3.2 實驗材料與試劑 25
3.3 實驗器材與設備 26
3.4 實驗方法 27
3.4.1 免疫微米磁珠之製備 27
3.4.2 標記生物素之奈米金粒子探針之製備 27
3.4.3 生物素檢測步驟 28
3.4.4 銀增強還原奈米金粒子探針 28
第四章 結果與討論 30
4.1生物素檢測實驗之最佳化條件 30
4.1.1 不同鹽濃度狀態下之最佳化測試 30
4.1.2 不同磁珠量捕捉奈米金探針之測試 31
4.1.3 奈米金探針與免疫磁珠結合時間的影響 32
4.1.4 樣品與磁珠結合時間之最佳化測試 34
4.2 奈米金探針檢測生物素與定量分析 35
4.3 銀還原增強奈米金探針訊號 37
第五章 結論 39
參考文獻 40
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論文全文使用權限:同意授權於2019-02-17起公開