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研究生： 吳愛麗
YUANA - ELLY AGUSTIN
論文名稱： 利用生物分子法的高產率表面測量
A BIOMOLECULE-BASED METHOD FOR HIGH THROUGHPUT SURFACE AREA MEASUREMENT
指導教授： 蔡伸隆
Shen-Long Tsai
口試委員: 李振綱
Cheng-Kang Lee
趙 玲
Ling Chao
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： 2014
畢業學年度： 102
語文別： 英文
論文頁數： 55
中文關鍵詞： 重组多肽 、纤维素结合结构域 、银纳米粒子 、表面面积测量
外文關鍵詞： recombinant peptide, cellulose binding domain, silver nanoparticles, surface area measurement
相關次數： 點閱：226  下載：3 
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    • 由於金屬奈米粒子的使用率漸漸增加，了解金屬奈米粒子的特性日漸重要，尤其是其表面區域，所以開發新的檢測方法便成為新的課題。本研究利用基因工程來達到檢測懸浮狀態之奈米粒子的表面之特性。此方法是將一種高選擇性的金屬結合肽將固定在纖維素之基材上，其用來抓取溶液中的金屬奈米粒子，然後再以被融合在金屬結合肽上的綠色螢光蛋白(Green Fluorescence Protein)來觀察螢光密度與奈米顆粒總表面積之關係。這個新的方法提供了快速且具選擇性的方法來檢測溶液中的特定金屬奈米粒子的表面積並且重組模組蛋白十分的穩定且可以儲存在攝氏4度下達六天。

    The increasing utilization of metallic nanoparticles in many areas of human activity has raised some concern to develop an effective and efficient method to understand the nature and characteristic of metallic nanoparticles, especially the surface area of metallic nanoparticles. In this study, an alternative approach for measuring the surface area of nanoparticles in suspension was studied with the aid of genetic engineering. A selective metal binding peptide that immobilized on a cellulose matrix was used to capture the metal nanoparticles from the solution and the Green Fluorescence Protein (GFP) that fused with metal binding peptide was used to investigate the correlation between fluorescence intensity with the total surface area of the nanoparticles. This newly method provides a fast and selective approach to detect the surface area of target metallic nanoparticles in a mixed solution. The recombinant modular protein quite stabile and can be used until 6 days in the 4oC storage temperature.

    摘要 iv ABSTRACT v ACKNOWLEDGEMENT vi TABLE OF CONTENT vii LIST OF TABLE ix LIST OF FIGURE x CHAPTER 1 12 1.1. Research Background 12 1.2. Objectives 13 CHAPTER 2 14 2.1. Metalic Nanoparticles 14 2.2. Surface Area Measurement 15 2.3. Recombinant Protein Production in Escherichia coli 17 2.4. Matrix for Protein Immobilization 19 2.5. Photosensing Using Green Fluorescence Protein 23 CHAPTER 3 26 3.1. Construction of Silver Binding Peptide 26 3.2. Construction of GFP-Ag4 27 3.3. Growth Media and Chemicals 27 3.4. Biosynthesis of Bacterial Cellulose Pellicles 28 3.5. Synthesis of Silver Nanoparticles 29 3.6. Immobilization of Silver Binding Peptide on Bacterial Cellulose 30 3.7. Analytic Performance 30 3.8. Long Term Storage Stability 31 3.9. Selective Fluorescence Enhancement Toward Over Other Nanoparticles 32 CHAPTER 4 33 4.1. Protein Expression and Purification of Silver Binding Proteins 33 4.2. Immobilization of Recombinant Protein on Bacterial Cellulose 37 4.3. Synthesis of Silver Nanoparticles by Chemical Reduction Process 38 4.4. Maximum Binding of Silver Nanoparticles to Recombinant Modular Protein (Ag4-CBD) onto Bacterial Cellulose Pellicles 40 4.5. Binding Analysis of Ag4-GFP with Silver Nanoparticles 41 4.6. Fluorescence Detection for Surface Area Measurement of Silver Nanoparticles using Ag4-GFP protein 42 4.7. Effect of Long Term Storage to the Stability of Recombinant Modular Protein 44 4.8. Selectivity of Modular Protein toward Nanoparticles 45 CHAPTER 5 47 REFERENCES 48 APPENDIX 51

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