溶膠-凝膠二氧化鈦型陣列生物感測器研發以及生醫應用

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研究生：	石慧美 Hui-mei Shih
論文名稱：	溶膠-凝膠二氧化鈦型陣列生物感測器研發以及生醫應用 Sol-Gel-Derived Titania Array-Based Biosensor for Biomedical Applications
指導教授：	董瑞安 Ruey-an Doong
口試委員:
學位類別：	碩士 Master
系所名稱：	原子科學院 - 生醫工程與環境科學系 Department of Biomedical Engineering and Environmental Sciences
論文出版年：	2005
畢業學年度：	93
語文別：	英文
論文頁數：	72
中文關鍵詞：	溶膠-凝膠、二氧化鈦、生醫應用
外文關鍵詞：	Sol-Gel, Titania, Biomedical
相關次數：	點閱：3 下載：0
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近年來，溶膠-凝膠的技術已廣泛地被應用於生物感測器的發展上。此外，陣列型生物感測器也在近年來感測器的研發上佔一席重要的地位。然而，由於金屬鈦的前趨物活性較大不易控制水解的速率，因此溶膠-凝膠二氧化鈦極少被應用於生物分子的包埋。而在本研究中，利用簡單的蒸汽沈降法，將尿素水解酵素，葡萄糖去氫酵素以及穀氨酸去氫酵素包埋於溶膠-凝膠二氧化鈦中以偵測尿素，葡萄糖以及麩氨酸。調控沈降靜置的時間以及溫度，生物分子能有效地包埋於溶膠凝膠的網狀結構中並保有其活性。由掃瞄式電子顯微鏡以及原子力顯微鏡圖像結果顯示當生物分子被包埋於二氧化鈦溶膠凝膠時，其溶膠凝膠材質的表棉結構及粗糙度大大地改變。而螢光顯微鏡的偵測結果可證明所製備得生物感測器能進行有效且精準的分析；對尿素的偵測其偵測極限可達20 □M（RSD=2.8%）；對葡萄糖的偵測其偵測極限可達50 □M（RSD=1.6%）；對麩氨酸的偵測其偵測極限可達78 □M（RSD=4.3%）。而對此三種分析物的偵測其偵測範圍可在2-3 層級之間，而對真實樣品的分析亦達良好的偵測效果。此外，利用此蒸汽沈降法以製備二氧化鈦溶膠凝膠薄膜其表面極微小的皸裂是可忽略的。總之，由研究結果可證明：利用簡單蒸汽靜置法，能將生物分子有效包埋於溶膠-凝膠二氧化鈦薄膜以製備陣列型生物感測器來作螢光的偵測以及應用於生醫用途。

The sol-gel-derived materials have recently become more attractive to immobilize biomolecules on the matrixes for biosensing application. In addition, array-based biosensor is a cutting-edge technology for the development for multi-analyte determination. However, titanium sol-gel material receives less attention on the immobilization as well as the biosensing application due to the high reactivity of the precursor. In this study, a simple vapor deposition method was developed to simultaneously immobilize urease, glucose dehydrogenase, and glutamate dehydrogenase on the glass matrix for the determination of urea, glucose and glutamate. Deposition time and temperature were optimized to control the transparency of TiO2 matrix which is suitable for optical detection. Results of SEM and AFM images showed that the doped enzymes into titania sol-gel film significantly changed the surface morphology. Urea, glucose, and glutamate could be efficiently detected by using urease-, GDH-, and GTDH-encapsulated array biosensors, and the detection limit was 20 □M for urea (RSD=2.8%), 50 □M for glucose (RSD=1.6%) and 78 □M for glutamate (RSD=4.3%). The array-based biosensors showed good analytical performance with dynamic range of 2-3 orders of magnitude. In addition, the responses of urea, glucose and glutamate to the array biosensors in real sample also showed well performances; the dynamic ranges were 1-100 □M, 40-10000 □M, and 80-10000 □M for urea, glucose, and glutamate, respectively. Also, the array biosensor showed a high relative enzyme activity and precise detection after 1-month storage. Furthermore, the shrinkage and crack of the TiO2 sol-gel matrix using vapor deposition method can be neglected. Results obtained in this study clearly demonstrate that this simple vapor deposition method can be successfully to form transparent titania sol-gel film for the fabrication of array-based biosensors that is suitable for optical detection of urea, glucose and glutamate for biomedical applications.

謝誌    Ⅰ
中文摘要    Ⅱ
Abstract    Ⅲ
Content Index    Ⅳ
Table Index    Ⅶ
Figure Index    Ⅷ
Chapter 1 Introduction    1
1-1 Motivation    1
1-2 Objectives    3
Chapter 2 Background and Theory    4
2-1 Biosensor    4
2-2 Enzyme-encapsulated biosensors    5
2-2-1 Glucose biosensors    5
2-2-2 Urea biosensors    7
2-2-3 Glutamate biosensors    7
2-3 Sol-Gel techniques    10
2-3-1 Introduction to sol-gel techniques    10
2-3-2 Titanium sol-gel techniques    15
2-4 Array-based biosensors    17
2-5 Fluorescence detections    18
Chapter 3 Materials and Methods    22
3-1 Materials    23
3-2 Preparation of array-based biosensor    24
3-3 Determination of enzyme activities    26
3-4 Substrate determinations    26
3-4-1 Solution form and real sample determination    28
3-4-2 Multi-analyte determinations    29
3-5 Fluorescence detections    30
3-6 Characterization of surface morphology    34
3-6-1 Atomic force microscopy (AFM)    34
3-6-2 Scanning electron microscopy (SEM)    35
Chapter 4 Results and Discussions    36
4-1 Fluorescence spectra of dye molecule in different pH values    36
4-2 Optimization of preparing array-based biosensor    39
4-2-1 Optimization of the components of enzyme solution    39
4-2-2 Optimization of the vapor deposition method    41
4-3 Surface morphology of the prepared titania sol-gel spots    42
4-4 Determination of enzyme activities    44
4-5 Optimization of biosensor performance    48
4-5-1 Urea determinations    48
4-5-1-1 Optimization of pH value    48
4-5-1-2 Optimization of buffer concentration    49
4-5-2 D-glucose determinations    52
4-5-3 L-glutamate determinations    56
4-6 Real sample determinations    59
4-7 Multi-analyte determinations    64
4-8 Long-term storage    66
Chapter 5 Conclusion    68
References    69

                                

1. Eggins, B. R. Chemical Sensors and Biosensors, 2002.
2. Yu, J. H.; Ju, H. X. Analytical Chemistry 2002, 74, 3579-3583.
3. Yu, J. H.; Liu, S. Q.; Ju, H. X. Biosensors & Bioelectronics 2003, 19, 401-409.
4. Yu, H. H.; Liu, S. Q.; Ju, H. X. Biosensors & Bioelectronics 2003, 19, 509-514.
5. Yu, J. H.; Ju, H. X. Analytica Chimica Acta 2003, 486, 209-216.
6. Clark, L. C. L. In Ann. Acad. Sci., 1962; Vol. 102, pp 29-33.
7. Altstein, M.; Bronshtein, A.; Glattstein, B.; Zeichner, A.; Tamiri, T.; Almog, J. Analytical Chemistry 2001, 73, 2461-2467.
8. H.Dislich Glastechn. Ber. 1971, 44, 1.
9. Chen, Q.; Kenausis, G. L.; Heller, A. Journal of the American Chemical Society 1998, 120, 4582-4585.
10. Pankratov, I.; Lev, O. Journal of Electroanalytical Chemistry 1995, 393, 35-41.
11. Wang, B.; Zhang, J.; Dong, S. Biosensors & Bioelectronics 2000, 15, 397-402.
12. Doong, R. A.; Tsai, H. C. Analytica Chimica Acta 2001, 434, 239-246.
13. Li, C. I.; Lin, Y. H.; Shih, C. L.; Tsaur, J. P.; Chau, L. K. Biosensors & Bioelectronics 2002, 17, 323-330.
14 McCain, K. S.; Harris, J. M. Analytical Chemistry 2003, 75, 3616-3624.
15. Besanger, T. R.; Chen, Y.; Deisingh, A. K.; Hodgson, R.; Jin, W.; Mayer, S.; Brook, M. A.; Brennan, J. D. Analytical Chemistry 2003, 75, 2382-2391.
16. Tsai, H. C.; Doong, R. A.; Chiang, H. C.; Chen, K. T. Analytica Chimica Acta 2003, 481, 75-84.
17. Wang, Q. L.; Lu, G. X.; Yang, B. J. Biosensors & Bioelectronics 2004, 19, 1269-1275.
18. Di, J. W.; Bi, S. P.; Zhang, M. Biosensors & Bioelectronics 2004, 19, 1479-1486.
19. Wang, J.; Pamidi, P. V. A.; Rogers, K. R. Analytical Chemistry 1998, 70, 1171-1175.
20. Wang, J. Analytica Chimica Acta 1999, 399, 21-27.
21. Wang, B. Q.; Li, B.; Deng, Q.; Dong, S. J. Analytical Chemistry 1998, 70, 3170-3174.
22. Liu, Z. J.; Deng, J. Q.; Li, D. Analytica Chimica Acta 2000, 407, 87-96.
23. Liu, Z. J.; Liu, B. H.; Kong, J. L.; Deng, J. Q. Analytical Chemistry 2000, 72, 4707-4712.
24. Du, D.; Yan, F.; Liu, S. L.; Ju, H. X. Journal of Immunological Methods 2003, 283, 67-75.
25. Chen, X.; Dong, S. J. Biosensors & Bioelectronics 2003, 18, 999-1004.
26. Zhang, Y.; He, P. L.; Hu, N. F. Electrochimica Acta 2004, 49, 1981-1988.
27. Yang, S. P.; Lu, Y. F.; Atanassov, P.; Wilkins, E.; Long, X. C. Talanta 1998, 47, 735-743.
28. Yao, T.; Takashima, K. Biosensors & Bioelectronics 1998, 13, 67-73.
29. Kunzelmann, U.; Bottcher, H. Sensors and Actuators B-Chemical 1997, 39, 222-228.
30. Li, J.; Chia, L. S.; Goh, N. K.; Tan, S. N. Journal of Electroanalytical Chemistry 1999, 460, 234-241.
31. Sampath, S.; Lev, O. Analytical Chemistry 1996, 68, 2015-2021.
32. Ramanathan, K.; Jonsson, B. R.; Danielsson, B. Analytica Chimica Acta 2001, 427, 1-10.
33. Wolfbeis, O. S.; Oehme, I.; Papkovskaya, N.; Klimant, I. Biosensors & Bioelectronics 2000, 15, 69-76.
34. de Marcos, S.; Galindo, J.; Sierra, J. F.; Galban, J.; Castillo, J. R. Sensors and Actuators B-Chemical 1999, 57, 227-232.
35. Lee, W. Y.; Kim, S. R.; Kim, T. H.; Lee, K. S.; Shin, M. C.; Park, J. K. Analytica Chimica Acta 2000, 404, 195-203.
36. Ogura, K.; Nakaoka, K.; Nakayama, M.; Kobayashi, M.; Fujii, A. Analytica Chimica Acta 1999, 384, 219-225.
37. Narang, U.; Prasad, P. N.; Bright, F. V.; Kumar, A.; Kumar, N. D.; Malhotra, B. D.; Kamalasanan, M. N.; Chandra, S. Chemistry of Materials 1994, 6, 1596-1598.
38. Imai, H.; Hirashima, H.; Awazu, K. Thin Solid Films 1999, 351, 91-94.
39. Matsumoto, T.; Murakami, Y.; Takasu, Y. Journal of Physical Chemistry B 2000, 104, 1916-1920.
40. Negishi, N.; Takeuchi, K. Thin Solid Films 2001, 392, 249-253.
41. Wang, H. L.; He, J. J.; Boschloo, G.; Lindstrom, H.; Hagfeldt, A.; Lindquist, S. E. Journal of Physical Chemistry B 2001, 105, 2529-2533.
42. Hanley, T. L.; Luca, V.; Pickering, I.; Howe, R. F. Journal of Physical Chemistry B 2002, 106, 1153-1160.
43. Areva, S.; Peltola, T.; Sailynoja, E.; Laajalehto, K.; Linden, M.; Rosenholm, J. B. Chemistry of Materials 2002, 14, 1614-1621.
44. Liu, H. M.; Yang, W. S.; Ma, Y.; Cao, Y.; Yao, J. N.; Zhang, J.; Hu, T. D. Langmuir 2003, 19, 3001-3005.
45. Hsu, J. P.; Nacu, A. Langmuir 2003, 19, 4448-4454.
46. Zhang, T.; Tian, B. Z.; Kong, J. L.; Yang, P. Y.; Liu, B. H. Analytica Chimica Acta 2003, 489, 199-206.
47. Moser, I.; Jobst, G.; Aschauer, E.; Svasek, P.; Varahram, M.; Urban, G.; Zanin, V. A.; Tjoutrina, G. Y.; Zharikova, A. V.; Berezov, T. T. Biosensors & Bioelectronics 1995, 10, 527-532.
48. G. Jobst, I. M., P. Svasek Sensor and Actuators B 1997, 43, 121-125.
49. Suzuki, M.; Akaguma, H. Sensors and Actuators B-Chemical 2000, 64, 136-141.
50. Perdomo, J.; Hinkers, H.; Sundermeier, C.; Seifert, W.; Morell, O. M.; Knoll, M. Biosensors & Bioelectronics 2000, 15, 515-522.
51. Palmisano, F.; Rizzi, R.; Centonze, D.; Zambonin, P. G. Biosensors & Bioelectronics 2000, 15, 531-539.
52. Moser, I.; Jobst, G.; Urban, G. A. Biosensors & Bioelectronics 2002, 17, 297-302.
53. Cho, E. J.; Bright, F. V. Analytical Chemistry 2001, 73, 3289-3293.
54. Cho, E. J.; Bright, F. V. Analytical Chemistry 2002, 74, 1462-1466.
55. Jobst, G.; Moser, I.; Varahram, M.; Svasek, P.; Aschauer, E.; Trajanoski, Z.; Wach, P.; Kotanko, P.; Skrabal, F.; Urban, G. Analytical Chemistry 1996, 68, 3173-3179.
56. Osborne, P. G.; Niwa, O.; Yamamoto, K. Analytical Chemistry 1998, 70, 1701-1706.
57. Tsai, H.-C. In Atomic Science; Nation Tsing Hua University, 2003; Vol. Ph.D, pp 103-139.

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