進階搜尋


 
系統識別號 U0026-0812200910174076
論文名稱(中文) 蛋白質晶片應用於酵素免疫分析法之研究
論文名稱(英文) Study of Protein Chips for Enzyme-Linked Immunosorbent Assay
校院名稱 成功大學
系所名稱(中) 工程科學系碩博士班
系所名稱(英) Department of Engineering Science
學年度 90
學期 2
出版年 91
研究生(中文) 侯曉晴
研究生(英文) Hsiao-Ching Ho
電子信箱 n9689419@ccmail.ncku.edu.tw
學號 n9689419
學位類別 碩士
語文別 中文
論文頁數 68頁
口試委員 口試委員-吳華林
指導教授-林裕城
口試委員-謝淑珠
中文關鍵字 微機電系統  蛋白質晶片  酵素免疫分析法 
英文關鍵字 ELISA  Protein Chip  MEMS 
學科別分類
中文摘要 本研究之目的,就是結合微機電技術設計及製作一種腫瘤檢驗晶片,在此晶片上進行酵素免疫分析法,作為危機評估、篩檢,進而評估病情,以期達到早期發現早期治療,減少癌症的死亡率。
研究的內容,主要分為兩方面:一方面是反應區晶片的設計及製作,包括比較不同晶片材質如:聚甲基丙烯酸甲酯、聚碳酸酯、聚苯乙烯及聚二甲基矽氧烷,反應區晶片表面之結構設計及反應區晶片之表面活化處理部分,探討上述之變因,進而設計出一個適合進行酵素免疫分析法之反應區晶片,並使用準分子雷射加工方式作為本研究中製作晶片之製程。
另一方面,則是設計一光學偵測系統,偵測因酵素免疫分析法所產生之化學冷光,主要架構是由光電倍增管將光源吸收並轉換成電壓訊號,透過鎖頻放大器作訊號處理,進而判斷在晶片上進行酵素免疫分析法之結果。
本研究之結果可證明在晶片上進行酵素免疫分析法是可行的,且晶片上反應具有良好之線性關係,在整個免疫分析處理的時間,相較於傳統方式縮短了許多,確實可以達到快速及準確的效果。

英文摘要 The goal of the study is to design and fabricate a tumor marker chip by using the techniques of MEMS. Apply enzyme-linked immunosorbent assay (ELISA) on the chip for the analysis of tumor marker in biological fluids. There are many advantages, including low reagent consumption, lowsample volume and waste production, faster response time, and feasibility to operate as a point-of-care testing.
The contents of the study have two parts. One is to design and fabricate the reaction area of the chip. The optimization of the chip was performed analyzing the parameters, including the materials that included PMMA, PC, PS and PDMS, the surface structures and the activation of chip’s surfaces.
The other is to develop a detection device for the chemiluminescence signal of ELISA reaction on chips. The signal is detection by an photomultiplier tube and processed by a lock-in amplifier.
The chip surface was activated for better reactivity and reproducibility after the surface coated with poly-L-lysine. Higher signal intensities were found on poly-L-lysine-treated chips. The results of the study show excellent sensitivity and linearity using the microchips for the immunobiochemical reaction.

論文目次 中文摘要 I
英文摘要 II
誌謝 III
目錄 IV
表目錄 VII
圖目錄 VIII
簡字及專有名詞 X
第一章 緒論 1
1.1 研究動機 1
1.2 生物晶片簡介 3
1.2.1 生物晶片的種類 5
1.2.2 生物晶片的應用 7
1.2.3 研發生物晶片的公司 9
1.3 研究架構 12
第二章 反應區晶片設計與製程技術 13
2.1反應區晶片設計原理 14
2.2 準分子雷射製程 15
2.2.1 準分子雷射加工機台 16
2.2.2 準分子雷射簡介 17
2.2.3 準分子雷射加工機制 19
2.3 光罩設計與製作 21
2.4 PMMA、PC、PS之製程 22
2.5 PDMS製程 23
第三章 光學偵測系統 24
3.1光學系統元件 25
3.1.1 光電倍增管 26
3.1.2 鎖頻放大器 28
3.1.3 切割器 31
3.2 光學偵測系統之架構 32
第四章 晶片反應區分析 33
4.1 晶片材質分析 35
4.2 晶片表面之活化處理 37
4.3 晶片結構分析 38
第五章 研究結果 42
5.1 晶片反應區材質、結構分析結果 42
5.2晶片反應區進行ELISA之實驗結果 49
5.3 光學偵測系統之實驗結果 52
第六章 結論與未來展望 57
6.1 結論 57
6.2 未來展望 59
參考文獻 60
附錄 A 63
附錄 B 65
自述 68

參考文獻 [1] K. Seiler, D.J. Harrison, A. Manz, “Planar Chips Technology for Miniaturization and Integration of Separation Techniques into Monitoring Systems,” J. Chromato., 593, pp. 253-258, 1992.
[2] J.S. Schultz, “Biotechnology-Based Recognition Elements,” IEEE Engineering in Medicine and Biology, pp. 210-216, March 1995.
[3] M. Washizu, T. Yamamoto, O. Kurosawa, N. Shimamoto, “Moleclar Surgery Based on Microsystems,” Conference on Solid-State Sensors and Actuators, pp. 473-476, June 1997.
[4] J. Stafford, J. Brignac, R. Gangadharan, M. McMahon, J. Denman, “Engineering in Genomics,” IEEE Engineering in Medicine and Biology, pp. 120-122, March 1999.
[5] B. LePioufle, P. Surbled, H. Nagia, K.S. Chun, Y. Murakami, E. Tamiya, H. Fujita, “Attachment of Cells on Microsystems:Application to the Gene Transfection,” Conference on Solid-State Sensors and Actuators, pp. 18-25, 1999.
[6] M.T. Cronin, R.V. Fucini, S.M. Kim, R.S. Masino, R.M. Wespi, C.G. Miyada, “Cystic Fibrosis Mutation Detection by Hybridization to Light-generated DNA Probe Arrays,” Hum. Mutat., pp. 244-255,1996.
[7] J. Michanel, D. Heller, “An Integrated Microelectronic Multiplex Hybridzation Systems : Design and Microfabrication,” IBC BioChip Technology, 1998.
[8] M.I. Song, K. Iwata, M. Yamada, K. Yokoyama, T. Takeuchi, “Multisample Analysis Using an Array of Microreactors for an Alternating-current Field-enhanced Latex Immunoassay,” Anal. Chem., 66, pp. 778-781, 1994.
[9] J. Briggs, P.R. Fanfili, “Quantitation of DNA and Protein Impurities in Biopharmaceuticals,” Anal. Chem., 63, pp. 850-859, 1991.
[10] A. Manz, D.J. Harrison, E. Verpoorte, H.M. Wildmer, “Planer Chips Technology for Miniaturization of Separation Systems : A Developing Perspective in Chemical Monitoring [Review]. In : P.R. Brown, E. Grushka, eds. Advances in Chromatography,” New York: Marcel Dekker, pp. 1-66, 1993.
[11] L.B. Koutny, D. Schmaizing, T.A. Tayor, M. Fuchs, “MicroChip Electrophoretic Immunoassay for Serum Cortisol,” Anal. Chem., 68, pp. 18-22, 1996.
[12] L.J. Kricka, O. Nozaki, S.Heyner, W.T. Garside, P. Wilding, “Applications of A Microfabricated Device for Evaluating Sperm Function,” Clin. Chem., 39, pp. 1944-1947, 1993.
[13] http://www.biochipmaster.com
[14] http://www.affymetrix.com
[15] http://www.nanogen.com
[16] http://www.hyseq.com
[17] http://www.biacore.com
[18] 國科會精密儀器中心儀器訓練手冊,2001。
[19] 準分子雷射微細加工機一般使用者訓練教材,2000/9。
[20] S.A. Berson, R.S. Yalow, “Assay of plasma insulin in human subjects by immunological methods,” Nature, 184, pp. 1648-1649, 1959.
[21] Roitt, Brostoff, Male原著,王聖予等人編譯,“免疫學”,藝軒出版社,中華民國八十五年十月第四版。
[22] D.S. Hage, “Immunoassay,” Anal. Chem., 71, pp. 294-304, 1999.
[23] J.P. Gosling, “A Decade of Development in Immunoassay Methodology,” Clin. Chem., 36, pp. 1408-1427, 1990.
[24] T.T. Ngo, “Developments in Immunoassay Technology,” Methods- ImmunoMethods, 22, pp. 1-3, 2000.
[25] M. Aizawa, “Immunosensor for Clinical Analysis,” Adv. Clin. Chem., 31, pp. 247-275, 1994.
[26] “Photomultiplier Tube Principle to Application,” HAMAMATSU, 1994.
[27] http://www.srsys.com
[28] “SR810 DSP Lock-in Amplifier Operating Manual and Programming Reference,” Stanford Research System Inc., 1993.
論文全文使用權限
  • 同意授權校內瀏覽/列印電子全文服務,於2002-07-05起公開。
  • 同意授權校外瀏覽/列印電子全文服務,於2002-07-05起公開。


  • 如您有疑問,請聯絡圖書館
    聯絡電話:(06)2757575#65773
    聯絡E-mail:etds@email.ncku.edu.tw