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標題: Investigation on piezoelectric quartz crystal biosensor in practical immunoassay
作者: 王智民
關鍵字: piezoelectric quartz crystal;壓電石英晶體;biosensor;生物感測器
出版社: 化學工程學系
Piezoelectric Quartz crystal has been used as transducer in biosensor research for years. Protein immobilization is the major concern of this quartz crystal microbalance (QCM) type of biosensor. In this research, different kind of physical and chemical protein immobilization methods were studied and compared with respect to their performance as well as the processes involved in the preparation.
Among chemical methods, the use of 11-mercaptoundecanol for gold electrode surface activation followed by carbodiimide condensation presents the bast responses when human serum albumin, HSA, used as insolubilized material for anti-HSA anti-HSA antibody detection. At anti-HSA concentration of 0.5mg/ml the change of frequency was 245Hz which was better than when the short chain mercaptans were used as activation agents.
Pyrrole based electro-induced polymerization was used for insolubilizing HSA on electrode surface. This novel immobilization approach in QCM study offers better sensitivities than the results from short chain mercaptans activation methods and is slightly inferior than the use of eleven-carbon mercaptan. The use of electro-induced polymerization to insolubilize protein on QCM electrode surface presents the advantages of simple, fast and easily scaled-up in its preparation which in turn favors the practical utilization of QCM in practical application.
In this study, the feasibility of using both sides of the quartz crystal for anti-HSA determination was examined. The results showed the responses were almost double when a flow cell design was uptaken which diverted the flow to other side after the sample flowed thru the first side of the electrode. The results indicate that double-side flow cell design offers not only the increase of sensitivity but also the possibility of determining dual analystes in one sample flow.

壓電晶體生物感測器是一種以質量變化做為感測依據的檢測方法。製作壓電生物感測器時,最主要的是如何固定蛋白質於晶體表面上。本論文中,主要探討物理性吸附法、化學共價鍵結法及電聚合高分子法等各種蛋白質固定法,進一步探討壓電晶體生物感測器在免疫分析上之應用。其中化學共價鍵固定法中的C11/EDC/HSA是利用長鏈11-mercaptoundecanol作表面活化,再以1-ethyl-3-(3-dimethylaminopropyl) carbodiimide作架接固定HSA的結果較佳,在檢測Anti-HSA濃度0.5mg/ml時頻率約為245Hz左右,不過其製作手續繁雜且時間需要較久。而電聚合pyrrole固定HSA的頻率變化約為210Hz左右,固定方法簡便又快速,應有實用於針對臨床診斷之潛力。
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