利用相位式表面電漿共振系統檢測免疫球蛋白鍵結之應用分析

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姓名

林萱(Shiuan Lin) 查詢紙本館藏

畢業系所

照明與顯示科技研究所

論文名稱

利用相位式表面電漿共振系統檢測免疫球蛋白鍵結之應用分析
(Immunoglobulin Binding Analysis by Utilizing a Surface Plasmon Resonance Phase-Measurement System)

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摘要(中)

本研究之動態式表面電漿共振感測器是以相位移干涉為基礎，以偵測器擷取光強訊號，代入五步相移法進行解相。由實驗結果可得知系統於3600秒下相位穩定度為1.2214度、系統靈敏度為1.1465✕10^4 (Degree/RIU)、系統折射率解析度可達1.0653✕10^-4 (RIU)。利用鹽水實驗可得折射率與相位差關係，進而去推算未知鹽水溶液折射率，經折射率計量測與實驗後內插所得之折射率，兩者誤差約為0.023 %。
於免疫球蛋白量測部分，利用濃度分別為200 μg/ml、100 μg/ml、50 μg/ml、25 μg/ml之Anti-IgG與固定於金膜上之IgG進行鍵結，所對應相位變化分別為：78.7476°、58.8533°、41.0506°、33.1399°，由相位與時間關係圖去計算結合速率常數k_a與解離速率常數k_d，其值分別為4.1139✕10^4
M^-1s^-1與2.8768✕10^-4s^-1。經由鹽水實驗與生物分子鍵結量測可得知SPR相位訊號隨待測液濃度改變之關係式，故可利用此關係曲線來進行未知濃度檢測。

摘要(英)

A dynamic surface-plasmon-resonance (SPR) sensor was developed based on interferometric phase measurements. Temporal intensity signals were acquired by photodetectors and processed by the Schwider-Hariharan five-step algorithm to obtain the phases. The achieved phase-detection stability was 1.2214 degrees in a 3600-second period and the system sensitivity was 1.1465×10^4 degree/RIU (refractive index unit). The corresponding system resolution was 1.0653×10^-4 RIU.
Salt-water mixture measurements were performed to characterize the relation between the measured phase difference and the refractive-index variation of the specimen. Several known concentrations of salt-water mixtures were flowed into the SPR system to measure the phases. A refractometer was utilized to measure the refractive indices. The results showed a linear correspondence between the phase difference and the refractive index. The measurement of an unknown salt-water mixture showed a relative refractive-index error of about 0.023 % between the refractometer measurement and the linear interpolation result from the SPR system.
For the antibody-antigen binding experiments, the phase differences caused by the injection of anti-IgG with concentrations of 25 μg/ml, 50 μg/ml, 100 μg/ml and 200 μg/ml were measured to be approximately 33.1399°, 41.0506°, 58.8533°, and 78.7476°, respectively. From the relation between the time and phase difference, the association rate constant ka and the dissociation rate constant kd were calculated to be 4.1139×10^4 M^-1s^-1 and 2.8768×10^-4 s^-1, respectively. By utilizing the relation between the SPR signals and the concentrations of the analyte from salt-water mixture measurements and antibody-antigen binding experiments, the concentration of unknown solutions can be estimated by this SPR system.

關鍵字(中)

★ 表面電漿共振
★ 生物分子

關鍵字(英)

★ Surface Plasmon Resonance
★ Immunoglobulin

論文目次

摘要i
Abstractii
誌謝iii
目錄iv
圖目錄vi
表目錄viii
第一章緒論1
1.1 前言1
1.2 研究動機2
1.3 文獻回顧2
1.3.1生物分子感測器-表面電漿感測器2
1.3.2 SPR免疫檢測技術5
1.3.3免疫球蛋白結構 6
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 激發表面電漿波條件14
2.2 Kretschmann組態下系統反射率15
2.2.1一層界面結構之系統反射率15
2.2.2二層界面結構之系統反射率16
2.2.3三層界面結構之系統反射率18
2.2.4四層界面結構之系統反射率19
2.3 動力學參數分析20
第三章光學解相理論 22
3.1 光學干涉原理22
3.2 相移干涉術24
第四章相位式表面電漿共振儀26
4.1 實驗系統架構26
4.2 系統解相方法30
4.3 感測片設計34
4.4 儀控程式38
4.5 訊號雜訊處理40
第五章實驗方法與結果分析41
5.1 實驗藥品41
5.2 藥品配製42
5.3 生物分子固定化程序43
5.4 SPR裝置44
5.5 實驗結果與模擬分析45
5.5.1實驗一 P極化光與S極化光量測45
5.5.2 實驗二系統穩定度與相位量測46
5.5.3實驗三鹽水實驗48
5.5.4 實驗四生物分子IgG與Anti-IgG結合量測51
第六章結論61
參考文獻62

參考文獻

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指導教授

陳怡君(Yi-Chun Chen)

審核日期

2012-7-31

推文