簡易檢索 / 詳目顯示

研究生: 葉怡均
YiChun Yeh
論文名稱: 醣包覆金奈米粒子與親醣蛋白間多價作用之研究
Multivalent Interactions Between Carbohydrate Encapsulated Gold Nanoparticles and lectins
指導教授: 陳家俊
Chen, Chia-Chun
學位類別: 博士
Doctor
系所名稱: 化學系
Department of Chemistry
論文出版年: 2003
畢業學年度: 91
語文別: 中文
論文頁數: 54
中文關鍵詞: 醣包覆金奈米粒子親醣蛋白多價作用陳家俊刀豆大腸桿菌
英文關鍵詞: Multivalent Interactions, Carbohydrate, Carbohydrate Encapsulated Gold Nanoparticles, lectins, FimH, Con A, E. coli
論文種類: 學術論文
相關次數: 點閱:227下載:0
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報
  • 自然界中許多生物體藉著多價反應機制來控制細胞表面受器與醣類分子間的辨識,這項機制為目前廣泛討論的課題。我們以醣類包覆金奈米粒子,研究其合成、性質及生物系統上的應用。此種金奈米粒子不僅擁有優異的水溶性,並且穩定存在各種離子濃度及酸鹼值的緩衝溶液中。我們首先以修飾甘露糖的金奈米粒子在電子顯微鏡下標定大腸桿菌特定的黏附蛋白,以及利用表面電漿共振光譜儀來偵測甘露糖、葡萄糖和半乳糖修飾之金奈米粒子與洋刀豆血球凝集素間的親合力及動力學解離常數。我們在此篇論文探討:以修飾醣分子的金奈米粒子做為高特異性之標定探針,及生物系統中多價鍵反應的優秀載體。

    Multivalent interactions between cell surface receptors and carbohydrates have been discovered in a number of biological processes. The synthesis, characterization and biological application of carbohydrate encapsulated gold nanoparticles (C-AuNP) are reported. C-AuNP is well dispersed and very stable without aggregation in the media of broad ion strength and pH ranges. We have demonstrated that mannose-encapsulated gold nanoparticles (m-AuNP) can be used as a probe to target mannose adhesin FimH of living bacterial type 1 pili using transmission electron microscopy, and explore the multivalent interactions between Con A and mannose-, glucose- and galactose-encapsulated gold nanoparticles studied using surface plasmon resonance (SPR). This work demonstrates that carbohydrate attached nanoparticles can be used as an efficient affinity label and a good a multi-ligand carrier in a biological system.

    總目錄………………………………………………….…………..I 中文摘要………………………………………………...…………Ⅳ 英文摘要…………………………………………………...………V 第一章、緒論……….………………………………………………1 1-1引言…….………………...……………………….……...……1. 1-2 奈米材料………………………………..………………..……2 1-2-1量子限量化效應………………………….……...……..…2 1-2-2表面電漿共振……….……...……………………………..3 1-2-3金奈米粒子之應用………………...……………………...4 1-3 多價效應………………………………………………….…...5 1-4 親醣蛋白與醣類分子作用力之量測…………………...…….7 1-4-1血球凝集抑制試驗………………………………………..8 1-4-2酵素連結醣蛋白分析法…………………………………..8 1-4-3等溫滴定卡計……………………………………………..9 1-4-4表面電漿共振偵測…………………………..……………10 第二章、實驗……………………………………………………….13 2-1研究動機與目的………..………………………...……………13 2-2以金奈米粒子標定醣蛋白……………………….……………16 2-2-1藥品……………..…………………………………………16 2-2-2儀器……………..…………………………………………17 2-2-3實驗步驟………..…………………………………………17 2-3 Concanavalin A及m-AuNP親合力之研究….………………..20 2-3-1藥品……………….…………………….…………………20 2-3-2儀器……………………….……………….………………20 2-3-3實驗步驟…………………….……………….…………..21 第三章、結果與討論…….………………………………………24 3-1 金奈米粒子標定FimH蛋白…………………………………..24 3-1-1合成金奈米粒子及標定FimH蛋白實驗參數…………..24 3-1-2修飾醣基之金奈米粒子(m-AuNP)醣量鑑定及性質……28 3-1-3 m-AuNP之離子穩定度……………………………………32 3-1-4 m-AuNP與free mannose對FimH蛋白競爭反應………34 3-2 量測Carbohydrate-AuNP與Con A親合力…………………...35 3-2-1量測Carbohydrate-AuNP與Con A親合力之實驗參數….35 3-2-2定量感應晶片表面醣基塗覆密度……………………….36 3-2-3測定抑制劑之抑制能力…………………………………36 3-2-4定量金奈米粒子醣包覆的比例與數量…………………47 第四章、結論……...………………………………………………49 未來展望…...………………………………………………………..50 參考文獻…………………………………………………………..51

    參考文獻
    [1] Brus, L. E.; Efros, A. L.; Itoh, T. J. Lumin. 1996, 354, 757.
    [2] Brus, L. E. J. Phem. Chem. 1994, 98, 3575.
    [3] Wang, Y. Z.; Qiao, G. W.; Liu, X. D.; Ding, B. Z.; Hu, Z. Q., Mater. Lett. 1993, 17, 152.
    [4] Rupp, J.; Birringer, R. Phys. Rev, 1987, B36, 7888.
    [5] Alivisatos, A. P. Science 1996, 271,933.
    [6] Chen, M.; Nikles, D. E. Nano Lett. 2002, 2, 211
    [7] Bawondi, M. G. J. Am. Chem. Soc. 1993, 115, 8709
    [8] Link, S.; El-Sayed, M. A. Int. Rev. Phys. Chem. 2000, 19, 409.
    [9] Chen, S. W.; Ingram, R. S.; Hostetler, M. J.; Pietron, J. J.; Murray, R. W.; Schaaff, T. G.; Khoury, J. T.; Alvarez, M. M.; Whetten, R. L. Science 1998, 280, 2098.
    [10] Link, S.; Beeby, A.; FitzGerald, S.; El-Sayed, M. A.; Schaaff, T. G.; Whetten, R. L. J. Phys. Chem. B 2002, 106, 3410.
    [11] Huang, T.; Murray, R. W. J. Phys. Chem. B 2001, 105, 12498.
    [12] Wilcoxon, J. P.; Martin, J. E.; Parsapour, F.; Wiedenman, B.; Kelley, D. F. J. Chem. Phys. 1998, 108, 9137-9143.
    [13] Mie, G. Ann. Physik 1908, 25, 377.
    [14] Genzel, L.; Marin, T. P.; Kreibig, U. Z. Physik 1975, B21, 339.
    [15] Hayat M. (Ed): Colloidal Gold: Principles, Methods and Applications. San Diego: Academic Press; 1 989.(2a)
    [16] Kiessling, L. L.; Pohl, N. L. Chem. Bio. 1996, 3, 71
    [17] Kiessling, L. L.; Gestwicki, J. E.; Strong, L. E. Curr. Opin. Chem. Biol.2000, 4, 696.
    [18] Kitov, P. I.; Shimizu, H.; Homans, S. W.; Bundle, D. R. J. Am. Chem. Soc. 2003, 125, 3284.
    [19] Sun, X.-L.; Kanie, Y.; Guo, C.-T.; Kanie, O.; Suzuki, Y.; Wong, C.-H. Eur. J. Org. 2000, 2643.
    [20] Choi, S. K.; Mammen, M.; Whitesides, G. M. J. Am. Chem. Soc. 1997, 119, 4103.
    [21] Roy, R.; Page, D.; Perez, S. F.; Bencomo, V. V. Glycoconjugate J. 1998, 15, 251.
    [22] Lin, C.-C.; Yeh, Y.-C.; Yang, C.-Y.; Chen, C.-L.; Chen, G.-F.; Chen, C.-C.; Wu, Y.-C. J. Am. Chem. Soc. 2002, 124, 3508
    [23] R.W. Wood, On a remarkable case of uneven distribution of light in a diffraction grating spectrum, Phil. Magm.1902, 4, 396.
    [24] E. Kretschmann, H. Raether, Radiative decay of non-radiative surface plasmons excited by light, Z. Naturforsch. 1968, 23A, 2135.
    [25] Kreuter, J. in Microcapsules and Nanoparticles in Medicine and Pharmacy; Donbrow, M., Ed.; CRC: Boca Raton, 1992.
    [26] R. Elghanian, J. J. Storhoff, R. C. Mucic, R. L. Letsinger, C. A. Mirkin, Science 1997, 277, 1078.
    [27] (a) de la Fuente, J. M.; Barrientos, A. G.; Rojas, T. C.; Rojo, J.; Cañada, J. ; Fernández, A.; Penadés, S. Angew. Chem. Int. Ed., 2001, 40, 2258. (b) Otsuka, H.; Akiyama, Y.; Nagasaki, Y.; Kataoka, K. J. Am. Chem. Soc. 2001, 123, 8226.
    [28] (a) Lee, Y. C.; Lee, R. T. Acc. Chem. Res., 1995, 28, 321. (b) Bertozzi, C. R.; Kiessling, L. L. Science, 2001, 291, 2357. (c) Mammen, M.; Choi, S.-K.; Whitesides, G. M. Angew. Chem. Int. Ed., 1998, 37, 2755.
    [29] Getwicki, J. E.; Strong, L. E.; Kiessling L. L. Angew. Chem. Int. Ed., 2000, 39, 4567.
    [30] Dimick, S. M.; Powell, S. C.; McMahon, S. A. Moothoo, D. N.; Naismith, J. H.; Toone, E. J. J. Am. Chem. Soc., 1999, 121, 10286.
    [31] Olsen, L. R.; Dessen, A.; Gupta, D.; Sabesan, S.; Sacchettini, J. C.; Brewer, C. F. Biochemistry, 1997, 36, 15073.
    [32] Mann, D. A.; Kanai, M.; Maly, D. J.; Kiessling, L. L. J. Am. Chem. Soc., 1998, 120, 10575.
    [33] (a) Gestwicki, J. E.; Cairo, C. W.; Strong, L. E.; Oetjen, K. A.; Kiessling, L. L. J. Am. Chem. Soc., 2002, 124, 14922. (b) Kiessling, L. L.; Strong, L. E.; Gestwicki, J. E. Annu. Rep. Med. Chem., 2000, 35, 321
    [34] Hernáiz, M. J.; de la Fuente, J. M.; Barrientos, Á. G.; Penadés. S. Angew. Chem. Int. Ed., 2002, 41, 1554.
    [35] Brust, M.; Walker, M.; Betthell, D.; Schiffrin, D. J.; Whyman, R. J. Chem. Soc. Chem. Commun., 1994, 801.
    [36] Kalinin, N. L.; Ward, L. D.; Winzor, D. J. Anal. Biochem., 1995, 228, 238. The RIP of each ligand of m-AuNP is calculated as: Ki (MeMan)/ [Ki (m-AuNP)the averaged number of mannose ligands on the m-AuNP].
    [37] Sato, T.; Ruth, R. Stabilization of Colloidal Dispersions by Polymer Adsorption; Surfactant Science Series, No.9; Marcel Dekker: New York 1980: pp 65.
    [38] Derewenda, Z.; Yariv, J.; Helliwell, J. R.; Kalb, A. J.; Dodson, E. J.; Papiz, M. Z.; Wan, T.; Campbell, J. EMBO J., 1989, 8, 2189.

    無法下載圖示
    QR CODE