研究生: |
TRAN NGUYEN THANH THUY TRAN - NGUYEN THANH THUY |
---|---|
論文名稱: |
以微白金導線為基礎所製備的電流式生物感測器進行丙氨酸轉氨酶之快速、經濟、且具選擇性的偵測 Micro-platinum Wire Based Amperometric Biosensors for Fast, Economic, and Selective Detection of Alanine Aminotransferase |
指導教授: |
曾婷芝
Tina T.-C. Tseng |
口試委員: |
Ming-Hua Ho
Ming-Hua Ho Chih-Ning Pao Chih-Ning Pao |
學位類別: |
碩士 Master |
系所名稱: |
工程學院 - 化學工程系 Department of Chemical Engineering |
論文出版年: | 2015 |
畢業學年度: | 103 |
語文別: | 英文 |
論文頁數: | 67 |
中文關鍵詞: | alanine aminotransferase 、biosensor 、microelectrode 、electrochemical 、ascorbic acid 、dopamine 、stability |
外文關鍵詞: | alanine aminotransferase, biosensor, microelectrode, electrochemical, ascorbic acid, dopamine, stability |
相關次數: | 點閱:273 下載:3 |
分享至: |
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最近電化學技術越來越備受關注,利用追蹤麩胺酸丙酮酸轉氨基酵素在血清裡的濃度上升,來診斷初期的肝臟疾病。而本研究期望製備出製程簡單、經濟效應性佳與高靈敏度的鉑絲線麩胺酸丙酮酸轉氨基酵素生物感測器,藉由比較兩種不同測試方式的麩胺酸丙酮酸轉氨基酵素感測探器並針對其優缺點進行探討。第一種是對電流(I)與麩胺酸丙酮酸轉氨基酵素活性作圖後直接由此圖中的斜率來進行分析(方法1),另一種則是在感測器得到麩胺酸丙酮酸轉氨基酵素訊號後再經過10分鐘才將其電流值與所對應的濃度值記錄下(方法2)。過氧化後的聚吡咯(PPy)薄膜與全氟磺酸薄膜(Nafion®)在本研究中是作為電極表面上的選擇性薄膜,用來抵擋干擾物所造成的訊號,增加感測器的專一性。這兩層薄膜雖然會降低過氧化氫(H2O2)的擴散速率使其較難到達電極表面,導致靈敏度下降,但卻可以有效的抵擋干擾物所產生訊號。由方法1所測試的麩胺酸丙酮酸轉氨基酵素感測器的響應時間為60秒,線性範圍為40-900 U/L,感測極限為15.92 U/L,靈敏度為1.908×10-5 nA/(s·U/L·mm2) (N = 10)。方法2的麩胺酸丙酮酸轉氨基酵素感測器的響應時間約為5秒,線性範圍為10-900 U/L,感測極限為8.48 U/L,靈敏度為0.059 nA/(U/L·mm2) (N = 10)。製備好的探針需要經過長時間操作與長期穩定度測試 (感測器須備存放在-20℃的環境中) (N = 5),同時兩種方式均要測試血清中麩胺酸丙酮酸轉氨基酵素的量來評估感測器的準確性。經過許多測試後可以得出以下結論:由方法1所製備的麩胺酸丙酮酸轉氨基酵素感測探針在高濃度的麩胺酸丙酮酸轉氨基酵素環境下結果較準確,同時也比較簡單來進行分析。相反地,方法2在低濃度的麩胺酸丙酮酸轉氨基酵素環境下較精準,同時具有較好的感測極限與線性範圍。
Lately, more attention has been drawn to electrochemical techniques for the early diagnosis of liver diseases by tracing the alanine aminotransferase (ALT) level in serum. In this study, a method for fabricating a fast, economic, and selective ALT biosensor based on the micro-platinum wire has been proposed and we compared two different methods for testing the ALT biosensor; –one was established by recording immediate slopes of current responses corresponding to different ALT activities (Method 1) and the other was established by recording 10 minute-delayed current responses corresponding to different ALT activities (Method 2). The permselective overoxidized polypyrrole and Nafion® were used for rejecting interferents. These layers added an additional barrier to lower the flux of H2O2 to the electrode surface; however, current noises from adding the interferents was successfully eliminated. ALT biosensors tested by Method 1 have response time of 60 sec, linear detection range of 40-900 U/L and sensitivity of 1.908×10-5 nA/(s·U/L·mm2) (N = 10) and that tested by Method 2 have response time of ~5 sec, linear detection range of 10-900 U/L, and sensitivity of 0.059 nA/(U/L·mm2) (N = 10). The operational, long-term and storage stabilities of ALT biosensors have been investigated (biosensors were stored at -20oC) (N = 5). ALT from spiked samples (ALT activities: 20, 200, 400, and 900 U/L) were also tested by both methods (N = 5). In general, Method 1 is more convenient to carry out and ALT biosensors tested by Method 1 can detect ALT at high concentration range (>300 U/L) more precisely, whereas ones tested by Method 2 have a better limit of detection, wider detection range and can detect ALT at low concentration range (<300 U/L) more precisely.
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