跳到主要內容

臺灣博碩士論文加值系統

(18.97.14.87) 您好!臺灣時間:2025/02/17 12:26
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:黃增煜
研究生(外文):Tseng-Yu Huang
論文名稱:肥大細胞類胰蛋白酶之電化學阻抗分析
論文名稱(外文):Detection of mast cell tryptase using electrochemical impedance spectroscopy
指導教授:陳志敏陳志敏引用關係吳嘉哲
口試委員:黃正昇沈靜慧
口試日期:2017-07-22
學位類別:碩士
校院名稱:國立中興大學
系所名稱:機械工程學系所
學門:工程學門
學類:機械工程學類
論文種類:學術論文
論文出版年:2017
畢業學年度:105
語文別:中文
論文頁數:56
中文關鍵詞:肥大細胞類胰蛋白酶金奈米半球陣列生醫晶片電化學阻抗分析
外文關鍵詞:mast celltryptasegold nano-hemisphere arraybiochipelectrochemical impedance spectroscopy
相關次數:
  • 被引用被引用:0
  • 點閱點閱:865
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
本實驗利用金奈米半球陣列生醫晶片量測肥大細胞類胰蛋白(tryptase)濃度。奈米半球具三維結構,可增加接附面積用以製成高靈敏度奈米生醫感測晶片。本實驗以電化學阻抗分析類胰蛋白濃度,並利用自我組裝層原理進行修飾。由於量化方式為電訊號,本實驗不需要使用二次螢光抗體標記,即可縮短檢測時間。相較於文獻中ImmunoCap的最低檢測極限約1ng/ml,本阻抗量測的最低檢測極限可以達到約1 pg/ml。本研究完成類胰蛋白濃度標準曲線,其中類胰蛋白阻抗是具有與濃度正相關之線性趨勢,R-squared為0.9828,將應用於臨床上麻醉科藥物過敏篩檢。由於過敏性休克往往屬於急性,會在極短的時間造成呼吸道腫脹,通氣困難而缺氧,使得手術中斷甚至造成致命可能。使用本實驗晶片對病患術前檢測是否對麻醉常用藥物過敏,應可降低無預期過敏性休克的發生率。同時,可將此生醫感測平台延伸至其他臨床藥物過敏的檢測。
The present study investigates a gold nano-hemisphere array biochip for measuring concentration of mast cell tryptase. Three-dimensional structure of the nano-hemisphere that increases the attachment area of gold surface can be used to fabricate a nano-medical sensor chip with high-sensitivity. In the present experiments, antibody and antigen in the biochip were modified by self-assembled monolayers. The sensitivity of the electrochemical impedance spectroscopy measurement reaches 1 pg/ml that is much more sensitive than the value of 1 μg/ml for ImmunoCap as reported in previous literature. The experiments show a linear relationship between the measured impedance and the tryptase concentration with a high R-squared value of 0.9828. The nano-hemisphere array biochip of present investigation can be used to detect allergic reactions in general and anaphylactic shock in particular. Anaphylactic shock causes respiratory tract swelling, hypoxia and hypotension, which could interrupt the operation and even lead to death of the patient. Development of the present biochip is able to avoid some specific allergens for the patients and reduce the risk of anaphylactic shock. At the same time, this bio-sensing platform can be extended to other clinical allergy testing.
摘要 i
Abstract ii
目錄 iii
圖表目錄 v
第一章、緒論 1
1.1研究動機與背景 1
1.2文獻回顧 3
1.2.1過敏現象 3
1.2.2 肥大細胞過敏反應 5
1.3研究目標 7
1.4論文架構 7
第二章、研究理論 8
2.1電化學 9
2.1.1電化學質傳現象 10
2.1.2電極雙層模型 11
2.1.3電化學電極量測 12
2.1.4電化學組抗分析法 14
2.2自我組裝層薄膜 21
2.2.1硫醇基鍵結於金基板 21
2.2.2 EDC與NHS錯合物修飾11-MUA 23
2.3抗體抗原專一性分析 24
2.3.1抗體抗原接合 24
第三章、實驗方法與材料 25
3.1抗體抗原調配 25
3.2奈米生醫檢測晶片 25
3.2.1奈米3D半球結構 26
3.2.2陽極氧化鋁膜背阻障層基板製作 27
3.2.3奈米半球AAO模具 28
3.2.4熱壓奈米半球PC試片 29
3.2.5奈米生醫感測晶片電極製備 30
3.2.6高溫退火 31
3.2.7奈米生醫感測晶片封裝 31
3.2.8電化學法沉積奈米金顆粒 32
3.3自我組裝薄膜 32
3.4接觸角 34
3.5抗體與抗原修飾 34
3.6 EC-LAB程式 35
第四章、結果與討論 36
4.1判定抗原專一性 36
4.2接觸角判斷MUA自我組裝層薄膜 37
4.3阻抗分析tryptase 38
4.3.1等效電路模型建立 38
4.3.2 Tryptase濃度標準曲線 42
4.3.3最低檢測極限 46
4.4電化學結果與現有檢測之比較 47
第五章、討論與未來展望 49
5.1結論 49
5.2未來展望 50
參考文獻 51
附錄 55
[1]Association, A. S., “2012 Alzheimer’s disease facts and figures,” Alzheimer's & Dementia, vol. 8, no. 2, pp. 131-168, 2012.
[2]Bard, A. J. and Faulkner, L. R.,“Fundamentals and applications,” Electrochemical Methods, vol. 2, 2001.
[3]Bellanti, J. A., “Literature review: the best new articles in the specialty of allergy, asthma, and immunology, 2004–2005,” in Allergy and Asthma Proceedings, 2006, vol. 27, no. 3, pp. 186-196: OceanSide Publications, Inc.
[4]Bird, T. D., “Genetic aspects of Alzheimer disease,” Genetics in Medicine, vol. 10, no. 4, pp. 231-239, 2008.
[5]Bogomolova, A., Komarova, E., Reber, K., Gerasimov, T., Yavuz, O., Bhatt, S., and Aldissi, M., “Challenges of electrochemical impedance spectroscopy in protein biosensing,” Analytical Chemistry, vol. 81, no. 10, pp. 3944-3949, 2009.
[6]Borrebaeck, C. A., Antibody engineering. Oxford University Press on Demand, 1995.
[7]Chaki, N. K. and Vijayamohanan, K., “Self-assembled monolayers as a tunable platform for biosensor applications,” Biosensors and Bioelectronics, vol. 17, no. 1, pp. 1-12, 2002.
[8]Chang, B.-Y. and Park, S.-M., “Electrochemical impedance spectroscopy,” Annual Review of Analytical Chemistry, vol. 3, pp. 207-229, 2010.
[9]Cheng, C., Peng, Y., Bai, J., Zhang, X., Liu, Y., Fan, X., Ning, B., and Gao, Z., “Rapid detection of Listeria monocytogenes in milk by self-assembled electrochemical immunosensor,” Sensors and Actuators B: Chemical, vol. 190, pp. 900-906, 2014.
[10]Ding, S.-J., Chang, B.-W., Wu, C.-C., Lai, M.-F., and Chang, H.-C., “Impedance spectral studies of self-assembly of alkanethiols with different chain lengths using different immobilization strategies on Au electrodes,” Analytica Chimica Acta, vol. 554, no. 1, pp. 43-51, 2005.
[11]Folkers, J. P., Laibinis, P. E., and Whitesides, G. M., “Self-assembled monolayers of alkanethiols on gold: comparisons of monolayers containing mixtures of short-and long-chain constituents with methyl and hydroxymethyl terminal groups,” Langmuir, vol. 8, no. 5, pp. 1330-1341, 1992.
[12]Foster, B., Schwartz, L. B., Devouassoux, G., Metcalfe, D. D., and Prussin, C., “Characterization of mast-cell tryptase-expressing peripheral blood cells as basophils,” Journal of Allergy and Clinical Immunology, vol. 109, no. 2, pp. 287-293, 2002.
[13]Goldstein, J., Newbury, D. E., Echlin, P., Joy, D. C., Romig Jr, A. D., Lyman, C. E., Fiori, C., and Lifshin, E., Scanning Electron Microscopy and X-ray Microanalysis: a text for biologists, materials scientists, and geologists. Springer Science & Business Media, 2012.
[14]Grieshaber, D., MacKenzie, R., Voeroes, J., and Reimhult, E., “Electrochemical biosensors-sensor principles and architectures,” Sensors, vol. 8, no. 3, pp. 1400-1458, 2008.
[15]Hamilton, R. G. and Oppenheimer, J., “Serological IgE analyses in the diagnostic algorithm for allergic disease,” The Journal of Allergy and Clinical Immunology: In Practice, vol. 3, no. 6, pp. 833-840, 2015.
[16]Harris, D. C., Quantitative chemical analysis. Macmillan, 2010.
[17]Johnson Jr, R. E. and Dettre, R. H., “Contact angle hysteresis. III. Study of an idealized heterogeneous surface,” Physical Chemistry, vol. 68, no. 7, pp. 1744-1750, 1964.
[18]Jorcin, J.-B., Orazem, M. E., Pébère, N., and Tribollet, B., “CPE analysis by local electrochemical impedance spectroscopy,” Electrochimica Acta, vol. 51, no. 8, pp. 1473-1479, 2006.
[19]Lai, K.-Y., Chang, C.-Y., Chou, T.-C., and Chang, H.-C., “Continuous monitoring of histamine release from single KU-812 cell with an electrochemical cellular chip,” in Nano/Molecular Medicine and Engineering (NANOMED), 2009 IEEE International Conference on, 2009, pp. 227-230: IEEE.
[20]Love, J. C., Estroff, L. A., Kriebel, J. K., Nuzzo, R. G., and Whitesides, G. M., “Self-assembled monolayers of thiolates on metals as a form of nanotechnology,” Chemical reviews, vol. 105, no. 4, pp. 1103-1170, 2005.
[21]Macdonald, J. R. and Barsoukov, E., “Impedance spectroscopy: theory, experiment, and applications,” History, vol. 1, no. 8, 2005.
[22]Retter, U. and Lohse, H., “Electrochemical impedance spectroscopy,” in Electroanalytical Methods: Springer, 2010, pp. 159-177.
[23]Sanz, M. L., Gamboa, P. M., and De Weck, A., “In vitro tests: basophil activation tests,” in Drug hypersensitivity: Karger Publishers, 2007, pp. 391-402.
[24]Schwartz, L., Yunginger, J., Miller, J., Bokhari, R., and Dull, D., “Time course of appearance and disappearance of human mast cell tryptase in the circulation after anaphylaxis,” Journal of Clinical Investigation, vol. 83, no. 5, p. 1551, 1989.
[25]Shan, X. C., Maeda, R., and Murakoshi, Y., “Micro hot embossing for replication of microstructures,” Japanese Journal of Applied Physics, vol. 42, no. 6S, p. 3859, 2003.
[26]Stephen, K., “Electrochemistry: Chemical reactions at an electrode, galvanic and electrolytic cells,” A Chem1 Reference Text, 2004.
[27]Sung, I.-H. and Kim, D.-E., “Surface damage characteristics of self-assembled monolayers of alkanethiols on metal surfaces,” Tribology Letters, vol. 17, no. 4, pp. 835-844, 2004.
[28]Toh, A., Wang, Z., and Wang, Z., “Ambient hot embossing of polycarbonate, poly-methyl methacrylate and cyclic olefin copolymer for microfluidic applications,” in Design, Test, Integration & Packaging of MEMS/MOEMS, 2009. MEMS/MOEMS'09. Symposium on, 2009, pp. 359-362: IEEE.
[29]Tsai, J.-J., Bau, I.-J., Chen, H.-T., Lin, Y.-T., and Wang, G.-J., “A novel nanostructured biosensor for the detection of the dust mite antigen Der p2,” International Journal of nanomedicine, vol. 6, p. 1201, 2011.
[30]Wang, J., “Nanomaterial-based electrochemical biosensors,” Analyst, vol. 130, no. 4, pp. 421-426, 2005.
[31]Warrington, R., Watson, W., Kim, H. L., and Antonetti, F. R., “An introduction to immunology and immunopathology,” Allergy, Asthma & Clinical Immunology, vol. 7, no. 1, p. S1, 2011.
[32]Wu, C.-C., Lin, C.-H., and Wang, W.-S., “Development of an enrofloxacin immunosensor based on label-free electrochemical impedance spectroscopy,” Talanta, vol. 79, no. 1, pp. 62-67, 2009.
[33]Yang, K.-L., Yiacoumi, S., and Tsouris, C., Electrical double-layer formation. CRC Press: Boca Raton, FL, 2004.
[34]Zhao, X.-Z. and Xu, X.-F., “The molecular fuzzy symmetry,” Acta Physicochimica Sinica, vol. 20, no. 10, pp. 1175-1178, 2004.
連結至畢業學校之論文網頁點我開啟連結
註: 此連結為研究生畢業學校所提供,不一定有電子全文可供下載,若連結有誤,請點選上方之〝勘誤回報〞功能,我們會盡快修正,謝謝!
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊