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研究生： 趙俞婷
Chao, Yu-Ting
論文名稱： 基於水凝膠介質中發光反應之可攜式生物傳感器開發
Development of a Portable Biosensor Based on Luminescent Reactions in Hydrogel Medium
指導教授： 帕偉鄂本
Urban, Pawel L.
口試委員: 曾建銘
Tseng, Chien-Ming
庫碼
Kumar, Vinoth
學位類別： 碩士
Master
系所名稱： 理學院 - 化學系
Department of Chemistry
論文出版年： 2021
畢業學年度： 109
語文別： 英文
論文頁數： 97
中文關鍵詞： 漂白水 、水凝膠 、物聯網 、肉新鮮度 、微控制器 、核甘酸 、農藥 、可攜式生物傳感器
外文關鍵詞： bleach, hydrogel, Internet of Things, meat freshness, microcontroller, necleotides, pesticides, portable biosensors
相關次數： 點閱：2  下載：0 
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    • 我們開發了一種可攜式無線生物傳感器—BioChemPen，用於快速分析吸附在固體表面上的物質。該裝置利用在水凝膠基質中發生的（生物）冷光反應。此裝置利用水凝膠作為化學感測器及使用光敏電阻來檢測光。光的產生是來源於當含有酶、電解質溶液和所有必需反應物的水凝膠接觸於含有目標分析物的固體樣品表面並將光敏電阻放置在靠近水凝膠的位置來辨識光的強度。我們使用MicroPython PyBoard微控制器板和其他低成本電子元件來控制BioChemPen而得到的結果會立即上傳到互聯網。我們將BioChemPen應用在檢測次氯酸鈉、三磷酸腺苷、二磷酸腺苷及有機磷農藥。其一我們利用了魯米諾和過氧化氫的水凝膠對含有次氯酸鹽的清潔劑的日常生活中常接觸的物體進行的分析。其二我們使用螢光素-螢光素酶-丙酮酸激酶水凝膠檢測肉類上的三磷酸腺苷和二磷酸腺苷來判斷肉的新鮮度。最後，我們運用了在乙醯膽鹼酯酶-膽鹼氧化酶-辣根過氧化物酶水凝膠檢測蔬菜中含有的有機磷農藥—甲基毒死蜱。另外，我們利用BioChemPen檢測了次氯酸鈉、三磷酸腺苷、二磷酸腺苷和甲基毒死蜱的偵測檢測分別為 7.95×10-11 mol mm-2、2.73×10-13 mol mm-2、2.35×10-12 mol mm-2、2.59×10-10 mol mm-2。

    We present BioChemPen – a portable wireless biosensor device for rapid analysis of substances adsorbed on solid surfaces. The device takes advantage of (bio)luminescent reactions taking place in a hydrogel matrix. In a typical embodiment, the active element of this device is a hydrogel disk (chemotransducer) containing enzyme(s), electrolyte solution, and all the necessary substrates. When the hydrogel is exposed to a solid sample surface containing the target analyte, light is produced. A photoresistor (phototransducer)—placed in close proximity to the hydrogel disk—detects the light. The BioChemPen operation is enabled by a MicroPython PyBoard microcontroller board and other low-cost electronic modules. The obtained results are immediately uploaded to the Internet cloud. In one application, we demonstrate analysis of hypochlorite-containing cleaning agents on daily use objects by an assay based on hydrogel embedded with luminol and hydrogen peroxide. In another application, we use luciferin-luciferase-pyruvate kinase hydrogel to detect adenosine triphosphate (ATP), and adenosine diphosphate (ADP) on meat to verify its freshness. Lastly, we demonstrate detection of organophosphate pesticides present on vegetables with the aid of acetylcholinesterase-choline oxidase-horseradish peroxidase hydrogel. The limits of detection for sodium hypochlorite, ATP, ADP, and chlorpyrifos-methyl (a pesticide) were 7.95×10-11 mol mm-2, 2.73×10-13 mol mm-2, 2.35×10-12 mol mm-2, and 2.59×10-10 mol mm-2, respectively.

    中文摘要 i Abstract ii 謝誌 iii Table of Contents iv List of Tables vi List of Figures vii List of Acronyms xvi Chapter 1: Introduction 1 1.1 Motivation 1 1.2 Luminescence 2 1.2.1 Chemiluminescence 3 1.2.2 Bioluminescence 5 1.2.3 Photoluminescence 5 1.2.3.1 Fluorescence 6 1.2.3.2 Phosphorescence 7 1.3 Biocatalysis 8 1.3.1 Enzymes 8 1.4 Single-point light sensor 10 1.4.1 Image sensor 13 1.4.1.1 Complementary metal oxide semiconductor 14 1.4.1.2 Charge coupled device 14 1.5 Three-dimensional printer 15 1.6 Microcontroller 16 1.7 Surface analysis 18 1.8 Internet-of-Things 18 1.9 Goals of this study 19 Chapter 2: BioChemPen for Rapid Analysis of Compounds Supported on Solid Surfaces 20 2.1 Introduction 20 2.2 Experimental section 24 2.2.1 Chemicals 24 2.2.2 Calculation of limit of detection and limit of quantitation 24 2.2.3 Design and construction of BioChemPen 25 2.2.4 Preparation of hydrogel disks 34 2.2.5 Data acquisition and treatment 36 2.2.6 Real samples used to demonstrate detection of bleach residues using gel-phase luminol chemiluminescent reaction 38 2.2.7 Real samples used to demonstrate detection of ATP using gel-phase luciferase bioluminescent reaction 39 2.2.8 Real samples used to demonstrate detection of a pesticide using gel-phase multi-enzyme chemiluminescent reaction 40 2.3 Results and discussions 40 2.3.1 Proof-of-concept: detection of bleach residues using gel-phase luminol chemiluminescent reaction 40 2.3.2 Detection of ATP, and evaluation of meat freshness using gel-phase luciferase bioluminescent reaction 50 2.3.3 Detection of ADP on surfaces using gel-phase luciferase bioluminescent reaction coupled with pyruvate kinase 56 2.3.4 Optimization of reaction conditions for the detection of pesticide (chlorpyrifos-methyl) using gel-phase multi-enzyme chemiluminescent reaction 56 2.3.5 Detection of a pesticide using gel-phase multienzyme chemiluminescent reaction 60 2.4 Conclusions 67 Chapter 3: Conclusions and Future Perspective 68 References 70 Appendix 1 91

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