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研究生:許遠玲
研究生(外文):Aline Natasia Kosasih
論文名稱:氧化石墨烯於紫膜晶片技術之應用
論文名稱(外文):Application of Graphene Oxide in Purple Membrane Chip Technology
指導教授:陳秀美陳秀美引用關係
指導教授(外文):Hsiu-Mei Chen
口試委員:陳秀美
口試日期:2012-07-25
學位類別:碩士
校院名稱:國立臺灣科技大學
系所名稱:化學工程系
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:英文
論文頁數:94
中文關鍵詞:氧化石墨烯紫膜晶片細菌視紫質
外文關鍵詞:graphene oxidepurple membrane chipbacteriorhodopsin
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此篇論文的目的在於探討如何結合氧化石墨烯(graphene oxide,GO)與紫膜(purple membrane,PM)以整合開發一種新穎的生物感測系統。GO是氧化態的石墨烯,其價位低廉也易於生產。而來自嗜鹽菌Halobacterium salinarum的PM則含有單一種蛋白質,細菌視紫質 (bacteriorhodopsin ,BR),為一種光驅動質子泵,具有獨特的光電性質。本研究開發一種簡易的方法將GO應用於PM生物晶片技術上,且並以原子力顯微鏡(AFM)、接觸角(contact angle)與光電流等分析來探討添加GO的成效。先製作結合GO與avidin的複合物(GO-avidin complex),然後當作架橋以將經biotin修飾的PM(b-PM)固定於以自排性單分子膜胺基化之 ITO表面。AFM分析的結果顯示,GO-avidin複合物的添加可增加PM晶片表面的平坦度,且GO用量的多寡對於平坦度影響十分重要。然而,GO添加對PM的光電流響應並沒有明顯的抑制效果。此外,也發現利用biotin與avidin作用的生物親和性固定化對於晶片上PM的塗覆率是很重要的,因為GO與PM間的非特異性吸附作用僅能產生較不穩定且微弱的光電流訊號。另一方面,僅單純將GO應用於PM晶片之層層塗覆製程:先以不同的塗覆方法將GO塗覆於晶片上,接著依序固定化avidin 與b-PM。接觸角分析証實每一種方法都可將GO塗覆上去,然而光電流也會因GO的結合而下降。在未來,此種整合GO的PM生物晶片可能開創一種獨特且功能較佳的生物分析技術。
The objective of this research was to explore the integration of graphene oxide (GO) with purple membrane (PM) in the approach for a novel bio-sensing system. As the oxidized form of graphene, GO possesses unique and stable structure derived from graphene. In addition, it is relatively cheap and easy to produce. PM from Halobacterium salinarum contains only one protein constituent, bacteriorhodopsin (BR) that acts as a light-driven proton pump providing novel photonic properties.
A facile way to introduce GO into PM chip technology was developed with evaluation of the GO addition effect by AFM, contact angle, and photocurrent analyses. GO-avidin complex was prepared and subsequently used as a linker to immobilize biotinylated PM onto ITO fabricated with an amine-terminated self-assembled monolayer. Morphological studies conducted with AFM revealed that GO addition through GO-avidin complex enhanced surface flatness of the PM chips and the amount of GO employed was critical to yield a distinguishable effect on the surface. Nevertheless, no significant inhibition of photoelectric responses generated from PM was observed on GO addition. The bioaffinity immobilization through biotin-avidin interaction was found to be important for high PM coverage, because the nonspecific binding directly between GO and PM yielded less stable and lower photoelectric responses. In addition, GO alone was integrated into the PM chip fabrication during the layer-by-layer assembly. GO was added with different coating methods prior to avidin and subsequently b-PM immobilization. The contact angle analysis confirmed the presence of coated GO in each method, while the chip photocurrents were reduced by GO integration. In the future, GO-integrated PM biochips might lead to a bioassay with unique and enhanced properties.
Abstract I
中文摘要 II
Acknowledgement III
Table of Contents IV
List of Figures VI
List of Tables XI

Chapter I. Introduction 1
Chapter II. Literature Review 3
2-1 Graphene and Graphene Oxide 3
2-1-1 Historical Background of Graphene and Grapene Oxide 3
2-1-2 Synthesis of Graphene Oxide 6
2-1-3 Morphological Structure of Graphene Oxide 8
2-1-4 Properties of Graphene Oxide 10
2-1-4-1 Dispersibility of Graphene Oxide 10
2-1-4-2 Electrical Properties of Graphene Oxide 12
2-1-5 Application of Graphene Oxide 13
2-2 Purple Membrane (PM) and Bacteriorhodopsin (BR) 15
2-2-1 Halobacterium salinarum 16
2-2-2 Structure and Properties of Purple Membrane (PM) 17
2-2-3 Bacteriorhodopsin Structure 19
2-2-4 Photocycle Mechanism of Bacteriorhodopsin 19
2-2-5 Photoelectric Response of Bacteriorhodopsin 23
2-2-6 Application of Bacteriorhodopsin 27
Chapter III. Experimental 29
3-1 Research Purpose 29
3-2 Materials and Equipments 30
3-2-1 Materials 30
3-2-2 Equipments 32
3-3 Procedure 32
3-3-1 ITO-NH2 Susbstrate Preparation 34
3-3-2 Buffers Preparation 35
3-3-3 GO-avidin complex preparation 35
3-3-3-1 GO-avidin complex prepared by covalent linking 35
3-3-3-2 GO-avidin complex prepared by biotin-avidin interaction 36
3-3-3-3 Purification of GO-avidin complex 36
3-3-4 PM functionalization 37
3-3-5 GO coating 38
3-3-5-1 GO coating by immersion method 38
3-3-5-2 GO coating by spin-coating 38
3-3-5-3 GO coating by fluid flow 39
3-3-6 PM photoelectric chip preparation 39
3-3-7 Analysis 40
3-3-7-1 Photocurrent Analysis 40
3-3-7-2 Contact Angle Measurement 41
3-3-7-3 Cyclic Voltammetry 41
3-3-8 Supplementary Experiment 42
Chapter IV. Result and Discussion 43
4-1 PM chip fabrication without GO 43
4-2 PM chip preparation with GO-avidin complex as the linker 49
4-2-1 Application of the complex of b-GO and activated avidin in PM-chip preparation 49
4-2-2 Application of the complex of GO and avidin in PM-chip preparation 57
4-3 Layer-by-layer PM chip preparation with GO 73
4-3-1 GO fabrication by the immersion method 74
4-3-2 GO fabrication by spin coating 80
4-3-3 GO fabrication by fluid flow 82
4-4 Reduction of GO 83
Chapter V. Conclusion and Future Work 88
5-1 Conclusion 88
5-2 Future Works 89
References 90
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