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標題: 奈米導電高分子於免疫層析檢測上的應用
Application of nano-conducting polymer to immunochromatographic test
作者: 吳芊彣
關鍵字: Immunochromatographic;Nano-conducting polymer
出版社: 化學工程學系
Biosensing system in association with immunochromatograph is considered to be an ideal technique which laypersons are capable to perform at any occasion on site. The most common example is the over-the-counter pregnancy test kit in drug stores.
In the research, it is our intention to utilize nanometer size conductive polymer beads as signal transduction marker to develop a novel conductometric immunosensor for field application. We use human serum albumin, HSA, its polyclonal antibody and poly-pyrrole as model system to explore the feasibility of this novel approach.
Poly-pyrrole appears to be superior than other conductive polymers for following reasons; 1. simple organic synthesis, 2. the conductivity can be easily manipulated by controlling oxireductive state of the beads and doping, 3. it is stable at ambient environment.
During the studies, it was found that the protein could hardly be immobilized onto the conductive polymer beads surface neither by ion-pair formation nor conductive polymer encapsulation during polymerization. A pyrrole derivative, pyrrole-2-carboxaldehyde is selected to copolymerized with pyrrole for the ease of forming stable conpolymer with an activated carboxy functional group for further protein immobilization. The protein thus insolubilized on the conductive polymer beads surface is stabilized and generates signal much greater than the previous methods.

本論文,是採用導電高分子 ( pyrrole、pyrrole-2-carboxaldehyde copolymer ) 為標誌物,以多株抗體 ( polyclonal anti-HSA ) 及人血清白蛋白抗原 ( HSA ) 為檢測模式蛋白,應用於免疫層析檢測 ( Immunochromatographic test, ICT ),分析其顯色強度與導電度進一步的比較及探討其檢測結果。免疫層析檢測適用於現場檢測 ( on-site testing ) ,不需經過長期訓練的操作者,在短時間內以單一步驟的檢測方式。目前一般商業化之標誌物多為膠體金與乳膠,導電高分子是一種較少見之標誌物,在過去極少被使用在免疫層析檢測上做為顯色及導電訊號傳遞的標誌物。
免疫層析檢測最重要的關鍵,在於標誌物的製備以及標誌物與分析物間的接合。本實驗一開始,採用聚吡咯為標誌物,聚吡咯與其他共軛導電高分子比較,聚吡咯則因 (1) 合成方法簡易,(2) 對水氣及空氣的穩定性佳,(3) 除了可經由氧化還原狀態調控其導電度之外,亦能經由質子酸摻雜 ( doping ) 來調控其導電度,所以具有相當大的應用潛力。
進而我們採用 pyrrole 衍生物,pyrrole — 2 - carboxaldehyde,在吡咯聚合的過程中,將pyrrole與pyrrole - 2 - carboxaldehyde一並加入,形成pyrrole - ( pyrrole-2-carboxaldehyde ) 共聚合物,pyrrole - 2 - carboxaldehyde 其結構上帶有醛基,氧化成為COOH,利用EDC與HSA共價鍵結縮合在一起。利用這種共價鍵結合的蛋白質固定化方法不僅製程較易控制,而且能提供穩定的訊號,供進一步的使用。
Appears in Collections:化學工程學系所

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