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研究生: 范仕遠
Fan, Shi-Yuan
論文名稱: 超高靈敏度的羧基改性氧化石墨烯之生醫感測於表面電漿子共振晶片之研發
Ultra Sensitivity of Carboxylated Graphene Oxide for Surface Plasmon Resonance Biosensors
指導教授: 邱南福
Chiu, Nan-Fu
學位類別: 碩士
Master
系所名稱: 光電工程研究所
Graduate Institute of Electro-Optical Engineering
論文出版年: 2017
畢業學年度: 105
語文別: 中文
論文頁數: 86
中文關鍵詞: 羧化氧化石墨烯羧基改性氧化石墨烯氧化石墨烯表面電漿子共振生物晶片生物感測分子動力學分析糖類抗原19-9
英文關鍵詞: Carboxylated graphite oxide, Biochip, Carbohydrate Antigen 19-9
DOI URL: https://doi.org/10.6345/NTNU202202070
論文種類: 學術論文
相關次數: 點閱:176下載:0
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  • 本論文使用以羧基改性氧化石墨烯為基材的表面電漿子共振感測晶片檢測牛血清蛋白以及糖類抗原19-9。首先使用牛血清蛋白比較羧基改性氧化石墨烯生醫晶片、氧化石墨烯生醫晶片、傳統生醫晶片的靈敏度。再由靈敏度最高的羧基改性氧化石墨烯生醫晶片檢測糖類抗原19-9。

    本論文也探討羧基改性氧化石墨烯、氧化石墨烯、石墨烯三種材料的導電性和單位折射率角位移靈敏度(S_RI)以及生物分子的吸附效率(E)比較。羧基改性氧化石墨烯擁有比氧化石墨烯更高的導電性,因為羧基能增加碳鏈的可饒性,且讓電荷載體更容易地沿著碳鏈移動。而導電性的提昇能增加表面電漿子極化的傳播常數,其與S_RI是正比的關係。表面電漿子共振系統被用來驗證羧基改性氧化石墨烯的S_RI度比氧化石墨烯更高。多功能電漿量測系統用來觀察羧基改性氧化石墨烯在640 mn-700 nm波長內表面電漿子共振角的特性。而穿透光譜系統以及掃描式電子顯微鏡則是被用來觀察羧基改性氧化石墨烯鍵結在晶片的特性。

    在實驗中證明了以羧基改性氧化石墨烯為基材的表面電漿子共振生物晶片於檢測牛血清蛋白時擁有超高的靈敏度和超低的檢測極限。0.275 mg/ml的羧基改性氧化石墨烯生醫晶片靈敏度比2 mg/ml的氧化石墨烯生醫晶片提升了2.1倍。除此之外,羧基改性氧化石墨烯生醫晶片在檢測牛血清蛋白抗體時最低檢測極可達10 fg/ml,明顯優於傳統生醫晶片。最後使用羧基改性氧化石墨烯進行糖類抗原19-9免疫反應的檢測。文獻指出有79%的胰腺癌病患體內糖類抗原19-9的含量在37 unit/ml以上。實驗結果發現以羧基改性氧化石墨烯為基材的表面電漿子共振生物晶片最低的抗原檢測極限可達10 unit/ml。

    基於本論文的實驗結果,以羧基改性氧化石墨烯為基材的表面電漿子共振生物晶片未來可以被使用在臨床診斷,廣泛應用於製藥行業、農業與環境檢測,大幅改善國人的健康品質。

    In this paper we proved that carboxyl-functionalized graphene oxide (GO-COOH) was used to detect pancreatic cancer indicator protein 19-9 with surface plasmon resonance (SPR) (Carbohydrate Antigen, CA19-9). We compare the sensitivity of GO-COOH biosensor、GO biosensor and traditional biosensor with bovine serum albumin. Next to detected CA19-9 by the highest sensitivity of GO-COOH biosensor.

    GO-COOH, graphene oxide (GO) and graphene were discussed with conductivity、SPR angular displacement sensitivity (S_RI) and adsorption efficiency of biomolecules. GO-COOH has better conductivity than GO because the carboxyl group is more flexible with carbon chain than other oxygen-containing functional groups and allows the charge carrier to move more easily along the carbon chain. The enhancement of conductivity increases the propagation constant of the surface plasma polarization, which is proportional to S_RI, and uses the SPR system to verify that GO-COOH is better than GO. Multifunctional plasmonic system (MPS) was been used to observe the characteristics of GO-COOH at the SPR angle of 640 mn-700 nm. The penetrating spectroscopy system and the Scanning electron microscope (SEM) were used to observe the properties of the GO-COOH bond on the chip.

    Experiment of SPR system is proved that GO-COOH-based surface plasmon resonance biochip has high sensitivity and low detection limit when detecting Anti-Bovine Albumin antibody (anti-BSA). 0.275 mg/ml GO-COOH biochip was 2.1 times better than the 2 mg/ml GO chip. In addition, the limit of detection of GO-COOH biochip low to 10 fg/ml in the detection of anti-BSA, significantly better than traditional biochip. Finally, the SPR system was used to detect the CA19-9 immune response. Previous literature indicates that 79% of patients with pancreatic cancer contains CA19-9 above 37 units/ml. The results show that the lowest antigen detection limit of GO-COOH as the biomaterial is 10 unit/ml.

    Based on the experimental results of this paper, the surface plasmon resonance biochip with GO-COOH as the biomaterial can be used in clinical diagnosis and widely used in the pharmaceutical industry, agriculture and environmental testing, and greatly improve the health quality of people.

    致謝 i 摘要 iii Abstract v 目錄 vii 圖表目錄 xii 第一章 緒論 1 1.1 前言 1 1.2 研究動機與目的 2 1.3 論文架構 3 第二章 基本原理與文獻回顧 4 2.1 表面電漿子共振 4 2.1.1 表面電漿子共振發展 4 2.1.2 表面電漿波 5 2.1.3 表面電漿子共振原理 5 2.1.4 表面電漿子共振的激發 8 2.2 石墨烯衍生物 10 2.2.1 石墨烯 10 2.2.2 氧化石墨烯 11 2.2.3 羧基改性氧化石墨烯 14 2.3 生物晶片原理與分子固定化 18 2.3.1 分子自組裝單層膜 18 2.3.2 分子固定化技術 20 2.4 等效介電常數 23 第三章 石墨烯衍生物基於表面電漿子共振感測器的靈敏度探討 25 3.1 表面電漿子共振感測器靈敏度的定義 25 3.1.1 折射率變化的靈敏度(S_RI) 25 3.1.2 生物分子吸附量的靈敏度(E) 26 3.2 石墨烯衍生物的導電性對於靈敏度的影響 27 3.2.1 導電性與折射率變化靈敏度的關係 27 3.2.2 導電性與介電常數和共振角變化的關係 27 3.3 羧基改性氧化石墨烯和氧化石墨烯的靈敏度比較 29 3.3.1 羧基改性氧化石墨烯增強導電性 29 3.3.2 羧基改性氧化石墨烯與氧化石墨烯的導電性比較 30 3.3.3 羧基能增加導電性的原因 31 第四章 實驗方法 33 4.1 實驗材料與設備 33 4.1.1 實驗材料 33 4.1.2 儀器設備 35 4.2 羧基改性氧化石墨烯的合成 36 4.3 表面電漿子共振感測晶片製作 39 4.4 驗證表面電漿子共振感測晶片製作完成 41 4.4.1 穿透光譜系統實驗 41 4.4.2 掃描電子顯微鏡(SEM)實驗 41 4.5 多功能電漿量測系統(MPS)實驗 41 4.6表面電漿子共振(SPR)微流道系統實驗 42 4.6.1 實驗目的 42 4.6.2 實驗溶液配置 43 4.6.3 微流道系統實驗介紹 44 4.6.4 系統流速最佳化 46 4.6.5 牛血清蛋白免疫分析 46 第五章 實驗結果與討論 48 5.1 驗證表面電漿子共振感測晶片製作完成 48 5.1.1 穿透光譜系統實驗結果 48 5.1.2 掃描電子顯微鏡(SEM)實驗結果 49 5.2 多功能電漿量測系統(MPS)實驗結果 50 5.3 表面電漿子共振(SPR)微流道系統實驗 52 5.3.1非特異性結合 52 5.3.2 分子動力學分析 53 5.3.3 羧基改性氧化石墨烯與氧化石墨烯的比較 55 5.3.4 羧基改性氧化石墨烯的低濃度檢測效果 61 5.4 不同晶片的標準差和變異係數統計分析 62 第六章 羧基改性氧化石墨烯透過表面電漿子共振技術檢測癌症指標糖類抗原199 64 6.1 糖類抗原199簡介 64 6.2 實驗目的 64 6.3 實驗方法 65 6.3.1 實驗溶液配製 65 6.3.2 表面電漿子共振(SPR)微流道系統實驗 66 6.4 實驗結果與討論 68 第七章 結論與未來展望 74 7.1 結論 74 7.2 未來展望 76 參考文獻 77

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