應用於生醫的微製作與網印電化學感測器;Microfabricated and Screen-printed Electrochemical Sensors for Biomedical Applications

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题名:	應用於生醫的微製作與網印電化學感測器;Microfabricated and Screen-printed Electrochemical Sensors for Biomedical Applications
作者:	蘇虹娜;Settu,Kalpana
贡献者:	電機工程學系
关键词:	大腸桿菌;生物傳感器;微細加工;絲網印刷;電化學;蛋白;免疫反應測定;Immunoassay;Biosensors;Microfabrication;Screen-printing;Electrochemical;Escherichia coli;Protein
日期:	2016-08-29
上传时间:	2016-10-13 14:37:58 (UTC+8)
出版者:	國立中央大學
摘要:	本研究發展電化學生物感測器，所使用的技術包括微製造和網印這兩種製造方法。這些感測器將應用在生物醫學上，如監控致病菌與檢測疾病的生物標記。本研究的微製造感測器是在玻璃基板上製造的指叉金微電極感測器，將用以檢測人體尿液中與牛奶樣品中的大腸桿菌。在細菌生長的期間，頻率範圍1 Hz到1 MHz的電化學阻抗譜測量，據以進行等效電路分析，從而推論出大腸桿菌的生長所引起電極表面之電雙層電容值的改變是阻抗變化的主因。在人體尿液檢測上，細菌生長期間由於會形成細菌生物膜累積在感測器表面上而產生阻抗變化。尿液中大腸桿菌的濃度可以靠著在細菌開始成長後的某一時間點測量在某一指定頻率之阻抗值然後對照在該時間點的校正曲線而得知。在牛奶檢測上，阻抗譜上可以觀察到阻抗值因大腸桿菌增生而降低的現象，這是由於牛奶中細菌代謝引起離子的濃度變化所致。建立阻抗降低10%所需的時間相對於細菌濃度的校正曲線，牛奶中大腸桿菌的濃度可以靠著測量10%阻抗變化的時間而得知。本研究的網印感測器是以碳為電極材料，將用於白蛋白與EN2蛋白的檢測。若用於檢測尿液中微量白蛋白，首先使用共價鍵固定法將人體白蛋白抗體固定在網印感測器表面，然後採用免疫反應測定原理，以計時電流法的電化學技術來對尿液中的白蛋白做定量檢測。為了要檢測尿液中的EN2蛋白，石墨烯被加入網印碳電極中以提高靈敏度；首先在網印石墨烯碳電極表面修飾接上能與EN2蛋白做特異性結合的單股DNA，然後採用循環伏安法來定量EN2蛋白的濃度。 ;This study focused on the development of electrochemical biosensors for biomedical applications, such as pathogenic bacteria monitoring and disease biomarker detection. Two methods were utilized for the fabrication of these sensors, namely the microfabrication and screen-printing techniques. The microfabricated biosensors were interdigitated gold microelectrode sensors designed and microfabricated on glass substrate for E. coli bacterial detection in human urine and cow milk samples. Electrochemical impedance spectroscopic measurements were carried out during bacterial growth over a frequency range of 1 Hz to 1 MHz. Based on equivalent circuit analysis, it was inferred that double layer capacitance was responsible for the impedance change caused by the E. coli growth. In human urine, the change in impedance during bacterial growth was due to formation and accumulation of bacterial biofilms on the sensor surface. The urinary E. coli concentration could be determined from the calibration curve at a specific frequency with a selected growth time. In cow milk, during E. coli growth, a decrease in impedance was observed due to the ionic concentration change in the milk caused by bacterial metabolism. The E. coli concentration in milk could be determined from the calibration curve of the bacterial concentration with respect to the time when the impedance changed by 10%. The screen-printed biosensors were carbon-based sensors for albumin and EN2 protein detection. For the detection of microalbumin in urine, anti-human albumin antibodies were immobilized on the screen-printed sensor surface by the covalent immobilization method. Then the immunoassay was achieved by quantitating the albumin in urine with the chronoamperometric (CA) electrochemical measurement technique. For the detection of EN2 protein in urine, graphene was incorporated in the screen-printed carbon electrode to enhance the detection sensitivity. The screen-printed carbongraphene sensor surface was modified with DNA strand which is specific to EN2 protein binding. Cyclic voltammetry measurement was employed for the quantitative detection of EN2 protein concentration.
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