Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/35510
標題: 非標定式阻抗分析免疫感測器於Enrofloxacin藥物的檢測
Label-Free Impedimetric Immuno-Sensor for the Detection of Enrofloxacin
作者: 林家鴻
Lin, Chia-Hung
關鍵字: enrofloxacin
enrofloxacin
immunosensor
electrochemical impedance spectroscopy
label-free
免疫感測器
電化學阻抗分析法
非標定式
出版社: 生物產業機電工程學系所
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摘要: Enrofloxacin是fluoroquinolones家族中最常被使用的抗菌劑之一,用來預防與治療家禽與畜類的疾病。研究中以11-mercapto-undecanoic acid (MUA)與金電極表面進行自組性單層膜之共架性鍵結,然後用EDC/NHS活化MUA尾端COOH官能基,使抗enrofloxacin抗體能被固定在MUA上以進行enrofloxacin檢測,電極修飾特性與免疫反應則在含有等量Fe(CN)6 3-/4-的磷酸緩衝溶液(PBS)中,以電化學阻抗分析法(electrochemical impedance spectroscopy, EIS)和循環伏安法(cyclic voltammetry, CV)進行分析。研究結果顯示最佳修飾條件為:(1)在37℃、15% RH的條件時MUA修飾最穩定;(2)在24小時內大部分的MUA可被EDC/NHS活化;(3)抗體在37℃、15%RH固定2小時最穩定;(4) bovine serum albumin (BSA)的阻隔效應在1小時內可達穩定;(5)利用Tween 20清洗10分鐘即可清除非特異性鍵結。在等效電路模擬上,一個電阻串聯兩個並聯電阻與電容(2R//C)之等效電路,可分辨電極與修飾膜間和修飾膜與溶液間的特性,而一個電阻串聯一個並聯電阻與電容(1R//C)之等效電路則具有分析簡單與低變異性等特點。研究結果也顯示,修飾條件的最佳化可使enrofloxacin檢測極限從10 ng/ml降低到1 pg/ml,此非標定式免疫感測器可以大幅簡化免疫檢測的步驟,並能提供足夠靈敏之檢測極限,將來可用於食品安全或臨床醫學低濃度生物分子的檢測。
Enrofloxacin, that is most extensive approval antibiotic in the fluoroquinolone family, is frequently used to treat and prevent the disease of food-producing animals. To immobilize the specific antibodt for enrofloxacin on an electrode can develop a lable-free immunosensor. The immunosensors were prepared by covalently binding anti-enrofloxacin antibodies onto a 11-mercapto-undecanoic acid (MUA) monolayer with the pretreatment of EDC/NHS activation on a gold electrode. Each modification process for the Au electrodes was investigated by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) in phosphate buffer saline(PBS) solution with Fe(CN)6 3-/4-. The optimal experiment conditions are : (1) MUA modification at 37℃ in 15% RH is more stable, (2) EDC/NHS can activate the most part of MUA after 24 h, (3)antibody immobilized at 37℃ in 15% RH presents stable after 2 h, (4) after 1 h, bovine serum albumin(BSA) blocking can finish, and(5) the nonspecific adsorption can be rinsed out for 10 min in Tween 20. In the equivalent circuit models, one resistor in series with two parallel RC circuits (2R//C) can clearly distinguish the impedance properties of the electrode-membrane and membrane-solution interface, and one resistor in series with one parallel circuit of resistor and capacitor (1R//C) possesses the advantage of simplification and stability of measurement. The optimal experiment conditions promote the limit of detection from 10 ng/ml to 1 pg/ml. The label-free impedance immunosensor supplies a sensitive and simple method for the enrofloxacin detection. In the future, the sensor can be applied to detect low concentration bio-molecule in food safety and clinical medical field.
URI: http://hdl.handle.net/11455/35510
其他識別: U0005-1308200821563100
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1308200821563100
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