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DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 鄭宗記 | |
dc.contributor.author | Hsin-Ju Chen | en |
dc.contributor.author | 陳欣儒 | zh_TW |
dc.date.accessioned | 2021-06-07T18:05:18Z | - |
dc.date.copyright | 2012-08-01 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-07-26 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16213 | - |
dc.description.abstract | 電容式生物感測器具有免標定的優點,具潛力發展成為手持式裝置應用於居家定點照護。傳統電容式感測器多半使用指插式電極,然而此種電極需要較繁複的微機電製程,以及具有電場不均勻分布的缺點。此外,利用電化學阻抗分析量測電容時,通常需要額外添加氧化還原物質以提高檢測靈敏度。因此,基於簡便之設計概念,本研究利用平行對電極以開發免標定與高靈敏度之電容式核酸適體生物感測器。此電極對為系統中的電容式傳感器,使用光碟片自製之金電極與氧化銦錫(indium tin oxide, ITO)薄膜作為對極,電極間以聚酯(PET)雙面膠帶貼合達到50 um之電極間距。於金電極上,藉由分子自組裝修飾核酸適體(aptamer)作為生物分子辨識元件,再修飾正十二烷基硫醇(1-dodecanethiol, DDT)以防止非特異性蛋白質吸附。此分子自組裝層的電化學特性顯示其亦能形成有效的絕緣層,適合用於開發電容式生物感測器。而本研究選擇凝血酶作為檢測標的物以驗證系統的有效性,使用自製電容式感測電路量測aptamer辨識到thrombin與其結合時產生之電容變化。在人體血清中,凝血酶檢測的濃度線性範圍為10-11 ~ 10-6 M,相關係數達0.97。於牛血清白蛋白(bovine serum albumin, BSA)與人血清白蛋白(human serum albumin, HSA)中,並無明顯的非特異性蛋白質吸附發生。因此,本研究成功利用電極對建構免標定電容式核酸適體生物感測器並應用於thrombin檢測。未來或許能透過置換不同的aptamer以檢測不同的目標蛋白質。 | zh_TW |
dc.description.abstract | Capacitive biosensors are label-free and promising for developing hand-held devices and point-of-care applications. However, conventional micro-fabricated capacitive sensors such as those based on interdigitated electrodes faced the challenges including delicate manufacture and non-uniform distribution of electric field. Furthermore, electrochemical impedance spectroscopy for capacitance measurements usually required extra addition of redox species to enhanced sensitivity. A label-free and sensitive capacitive aptasensor was developed with simple design based on electrode pairs. The electrode pair was the transducer of the capacitive aptasensor, which was composed of a gold electrode (from compact discs) and an indium tin oxide (ITO) film with micrometer separation by a double-side PET tape. On the gold electrode was layered with aptamers as the bio-recognition elements and then a self-assembled monolayer of 1-dodecanethiol to prevent non-specific protein binding. Electrochemical characterization proved that the self-assembly monolayer also formed effective insulating layer adequate for capacitive sensing. Thrombin was selected as a model analyte to validate the assay. The capacitance change originated from thrombin binding with aptamers was measured with a home-made capacitance sensing circuit by switched capacitor technology. The linear range for thrombin detection in human serum was from 10-11 to 10-6 M with a regression coefficient of 0.97. Non-specific binding of bovine serum albumin (BSA) and human serum albumin (BSA) were not observed in the proposed aptasensor. This work demonstrated the successful application of label-free capacitive aptasensor for thrombin detection. It could be applied in the detection of other target proteins by replacing different aptamers. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T18:05:18Z (GMT). No. of bitstreams: 1 ntu-101-R99631005-1.pdf: 15395094 bytes, checksum: 7069e06b479537f63701d7f6e900c197 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 口試委員審定書 i
致謝 ii 中文摘要 iii Abstract iv Table of Contents v List of Tables vii List of Figures viii Chapter 1 Introduction 1 Chapter 2 Literatures Review 5 2.1 Capacitive biosensors 5 2.1.1 Electrode types 6 2.1.1.1 Interdigitated electrodes 6 2.1.1.2 Electrodes based on the electrode/solution interface 8 2.1.2 Sensing methods of capacitive biosensors 8 2.1.2.1 Impedance measurements 9 2.1.2.2 Potentiostatic step measurements 10 2.1.2.3 Switched capacitor technology 11 2.2 Self-assembled monolayers 11 2.2.1 Formation process of self-assembly monolayers 12 2.2.2 General aspects of thiol SAMs on gold 13 2.2.3 Advantages of self-assembled monolayers based on thiols 14 2.2.4 The electrical impact of SAMs on material properties 15 2.3 Compact discs 17 2.3.1 Structure of compact discs 17 2.3.2 Surface chemistry on CD-Rs for self-assembled monolayers 17 2.3.3 CDtrodes for electroanalytical chemistry 18 2.4 Aptamer 19 2.4.1 Synthesis of aptamer 20 2.4.2 Thrombin-binding aptamer 22 2.4.3 Applications of aptamer 23 2.5 Thrombin 27 2.5.1 Structure of thrombin 27 2.5.2 Biological functions of thrombin in blood coagulation 29 2.5.3 Biological functions of thrombin in diseases 30 2.5.4 Diagnosis methods of thrombin generation 31 Chapter 3 Materials and Methods 33 3.1 Chemicals and materials 33 3.2 Apparatus 34 3.3 Aptamer 34 3.4 Preparation of gold electrodes from compact discs 34 3.5 Characterization of the CDtrodes by cyclic voltammetry 34 3.6 Atomic force microscopy (AFM) analysis of gold substrates 35 3.7 Fabrication of aptamer-modified gold CDtrodes 35 3.7.1 Characterization of the modified CDtrodes by cyclic voltammetry 35 3.7.2 Characterization of the modified CDtrodes by electrochemical impedance spectroscopy 36 3.8 Capacitive measurements of thrombin 36 Chapter 4 Results and Discussion 37 4.1 Electrochemical behavior of the CDtrodes 37 4.2 Characterization of the morphology of the gold CDtrodes 37 4.2.1 Analysis by atomic force microscopy (AFM) 37 4.2.2 Comparing with other commercial gold electrodes 37 4.3 Effects of different modified CDtrodes on electrochemical characters 38 4.3.1 Cyclic voltammetry of the modified CDtrodes 38 4.3.2 Electrochemical impedance spectroscopy of the modified CDtrodes 39 4.4 Feasibility of capacitive aptasensor 41 4.5 Specificity of the capacitive aptasensor for thrombin detection 41 4.6 Determination of thrombin 42 Chapter 5 Conclusions 44 References 45 | |
dc.language.iso | en | |
dc.title | 免標定電容式核酸適體生物感測器運用於凝血酶的檢測 | zh_TW |
dc.title | Label-Free Capacitive Aptasensor for Thrombin | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳力騏,吳靖宙 | |
dc.subject.keyword | 電容式,電極對,核酸適體,分子自組裝,凝血酶, | zh_TW |
dc.subject.keyword | Capacitive,Electrode pair,Aptamer,Self-assembly monolayer,thrombin, | en |
dc.relation.page | 85 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2012-07-26 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 生物產業機電工程學研究所 | zh_TW |
顯示於系所單位: | 生物機電工程學系 |
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