利用光纖式表面電漿共振免疫感測器於退伍軍人菌之檢測

Application of Side-Polished Fiber Immunosensor Based on Surface Plasmon Resonance for the Detection of legionella pneumophila

楊伊雯(Yi-Wen Yang)

指導教授 ： 顏聰榮 ； 蔡五湖

大同大學/工程學院/生物工程學系所/碩士(2006年)

摘要

過去十年來，表面電漿共振(SPR)已經被證明成為生物分子間相互作用的強而有力和定量之探查工具。SPR的優勢在於高靈敏性且不需任何螢光或其他標的物之相互作用物。本研究是利用光纖式表面電漿共振免疫感測器於退伍軍人菌檢測之研發與應用。調查三種固定抗退伍軍人菌之單株抗體的方法，分別為(1) protein A 吸附層、(2) self-assembled 11-MUA層、(3) self-assembled 11-MUA與protein A層。利用掃描電子顯微鏡(SEM)與原子力顯微鏡(AFM)觀察方法(1)與(3)表面protein A吸附量，結果顯示方法(3)是利用化學鍵結的方式固定protein A，所以比物理吸附protein A有較佳的吸附量。利用氣態鹵素光式SPR免疫感測器分析，方法(1)與(2)只有少量的抗體被固定，並無法有明顯的SPR效應產生；然而方法(3)有明顯SPR效應的產生，所以為最佳固定抗退伍軍人菌之單株抗體的方法。以最佳的固定方法(3)做敏感度試驗，氣態鹵素光式SPR免疫感測器之偵測極限為101 CFU/ml；流體雷射式SPR免疫感測器之偵測極限為102 CFU/ml。光纖式表面電漿共振免疫感測器可用來檢測退伍軍人菌且有高敏感度的偵測極限。

關鍵字

表面電漿共振 ； 免疫感測器 ； 退伍軍人菌

並列摘要

Surface plasmon resonance (SPR) sensing has been demonstrated in the past decade to be an exceedingly powerful and quantitative probe of the interactions of a variety of biopolymers. An advantage of SPR is its high sensitivity without any fluorescent or other labeling of the interactants. An immunosensor using side-polished optical fiber based on surface plasmon resonance was proposed for the detection of Legionella pneumophila. Three immobilization strategies of Mab against L. pneumophila were investigated, namely, (1) protein A adsorption layer, (2) self-assembled 11-MUA layer, and (3) self-assembled 11-MUA and protein A layer. Using scanning electron microscope (SEM) and atomic force microscope (AFM), the protein A absorbed on surfaces in method (1) and (3) were observed. The result showed that the bound protein A by chemical bond in method (3) was better than it by physical absorption. From the analysis of SPR spectrum with halogen light in dip-and-dry method, there were only few Mab against L. pneumophila immoblilized by the method (1) and (2) whose wavelength shifts of SPR were unobvious. However, the SPR effect was obvious by the method (3), so the self-assembled 11-MUA with protein A layer was the best immobilization method of Mab against L. pneumophila comparing to the other two methods. The immnuosensor based on SPR spectrometer with halogen light in dip-and-dry method for the detection of Legionella pneumophila had the detection limit of 101 CFU/ml. The detection limit of an immnuosensor based on SPR powermeter with laser source with flow-through method was 102 CFU/ml. Hence, the side-polished optical fiber immunosensor based on surface plasmon resonance are high sensitivity and able to detect the Legionella pneumophila.

並列關鍵字

Surface Plasmon Resonance ； Immunosensor ； Legionella

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