博碩士論文 101226047 詳細資訊




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姓名 張綺珊(Chi-Shan Chang)  查詢紙本館藏   畢業系所 光電科學與工程學系
論文名稱 結合表面電漿共振及溫度控制於免疫球蛋白鍵結之檢測分析
(Immunoglobulin Binding Analysis by Utilizing a Surface Plasmon Resonance Phase-Measurement System with Temperature Control)
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摘要(中) 大多生物分子在各種溫度下會有不同的活性反應,故本研究以干涉式相位量測的表面電漿感測器為基礎,加入溫度控制裝置,進行P極化光、不同濃度的鹽水實驗及免疫球蛋白IgG與Anti-IgG之鍵結實驗,並用五步演算法進行解相分析。以純水及鹽水實驗量測系統表現,得到的系統穩定度為0.4667度,系統靈敏度為1.617x10^3 (Degree/RIU),系統折射率解析度可達2.886x10^-4 (RIU)。

本研究將IgG附著於金膜上,通入Anti-IgG進行鍵結,並討論在常溫下及有使用溫度控制,亦即生物體溫度(36 )下的實驗情況,在有使用溫度控制下的IgG及Anti-IgG之結合所產生的相位變化為6.72度,未使用溫度控制的相位變化為3.04度,兩者差異為3.68度。使用溫度控制裝置將溫度固定於實際生物分子的生存溫度時,將能從實驗過程中得到更接近於現實的生物分子活性反應,因此結合溫度控制裝置於表面電漿共振系統,將適用於進行更多種類的生物分子檢測。

摘要(英) Most biomolecules have different reaction activities at various temperatures. In this study, a temperature control apparatus was incorporated into a surface plasmon resonance (SPR) sensor, which is based on the interferometric phase measurements. Three kinds of experiments were performed, including P-polarized light measurements, salt-water mixture measurements and antibody-antigen binding experiments. The acquired temporal intensity signals were processed by the Schwider-Hariharan five-step phase-shifting algorithm to obtain the phases. The achieved phase-detection stability was 0.4667 degrees, and the system sensitivity was 1.617x10^3 degrees/RIU (refractive index unit). In addition, the corresponding system sensitivity was 2.886x10^-4 RIU.

In the third experiment, IgG was first binded on the Au film, and then anti-IgG was pumped into the flow cell to bind with IgG. The experiments were performed at two temperatures: at room temperature and at 36 degrees Celsius. The phase changes during the binding process were 6.72 degrees at 36 degrees Celsius, and 3.04 degrees at room temperature. The difference in phase changes between the two temperatures was 3.68 degrees. By using temperature control, the experiment results would be closer to the reality, which enables more varieties of biomolecular reactions to be detected by this SPR system.

關鍵字(中) ★ 表面電漿共振
★ 溫度控制
關鍵字(英)
論文目次 摘要 I

ABSTRACT II

誌謝 IV

目錄 V

圖目錄 VIII

表目錄 XII

第一章 緒論 1

1.1前言 1

1.2文獻回顧 1

1.2.1生物分子感測器-表面電漿感測器 1

1.2.2 SPR免疫檢測技術 4

1.2.3 免疫球蛋白結構 6

1.3 研究動機與目的 7

1.4 論文架構 7

第二章 基本理論 8

2.1 表面電漿原理 8

2.1.1 表面電漿波之色散關係式 8

2.1.2 SPR電漿波的產生條件 12

2.2 KRETSCHMANN組態下系統反射係數 13

2.2.1 一層介面結構之系統反射係數 13

2.2.2 二層介面結構之系統反射係數 16

2.2.3 三層介面結構之系統反射係數 17

2.2.4 四層介面結構之系統反射係數 19

第三章 相位式表面電漿共振儀 20

3.1 系統架構 20

3.2 系統解相方法 25

3.2.1 光學干涉原理 25

3.2.2 相移干涉術 27

3.3 感測片設計 30

3.4 儀控程式 32

第四章 實驗方法 42

4.1 實驗藥品 42

4.2 藥品配製 43

4.3 生物分子固定化程序 43

4.4 實驗程序 44

第五章 實驗結果與分析 46

5.1 P極化光的量測 46

5.2 系統穩定度與相位量測 48

5.3 鹽水實驗 49

5.4 生物分子IGG與ANTI-IGG結合量測 51

5.4.1 常溫下量測 51

5.4.2 生物體溫下( C)量測 54

第六章 結論 58

參考文獻 59

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Available: http://refractiveindex.info/?group=LIQUIDS&material=Water.

指導教授 陳怡君 審核日期 2015-8-31
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