纳米材料的制备及其在生物传感和检测中的应用
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摘要
当物质的尺寸下降到纳米尺度(1-100nm)时,其量子限域效应,表面与界面效应,小尺寸效应,宏观量子效应将令纳米材料展现出许多独特的性质,在电子,材料,光学,生物等领域都具有广泛的应用前景。近年来,纳米材料在生物传感和生物检测方面的应用越来越受到世人瞩目。本论文制备了基于碳纳米管和氧化锌协同作用的生物传感器并测定其特性；利用超声辅助的方法,合成具有良好生物相容性的量子点；将量子点标记到载有药物的聚合物上,在荧光显微镜下观察聚合物在动物体内的分布,并分析其代谢情况。主要研究内容如下：
1由于碳纳米管对过氧化氢的催化性以及氧化锌的高等电位,制备了基于此两种材料协同作用的葡萄糖传感器。纳米材料的应用,极大的提高了传感器的性能,该传感器为生物活性酶提供了一个良好的微环境并有效地防止酶泄漏,能检测溶液中葡萄糖的存在并具有较高的灵敏度、抗干扰性,及其较好的重复性。
2以超声辅助的方法,水相制备了CdSe和CdSe/CdS量子点,该量子点具有较好的水溶性,通过接枝巯基乙酸(MAA)使量子点具备较好的生物相容性,能够与常见生物分子如抗体,蛋白,DNA等相连,在免疫荧光分析,生物分子标记方面有很大的应用价值。
3将CdSe/ZnS量子点标记到载有药物的聚合物分子Poly-PTX上,用粒径仪,TEM等研究粒子的大小及形貌,将标记了量子点的聚合物注射到裸鼠体内,制作裸鼠各组织的冰冻切片,利用高分辨率荧光显微镜观察聚合物分子在裸鼠体内各个组织的分布,对Poly-PTX做一个初步的靶向分析,并分析其在裸鼠体内的代谢情况,实验结果表明量子点是一种高效、低毒、稳定的荧光标记物。本实验对量子点在生物成像领域的应用作了初步的探索。
When the size of some substances decrease to nanometre scale, they will show many special properties owing to their quantum confinement effect, surface and interface effect, small size effect and macroscopic quantum effect, which will be widely used in electronic science, material science, optics and biology. Recently, nanomaterials are more and more attractive because of their application in biosensor and biodetection. In this thesis, a biosensor based on synergistic action of carbon nanotubes and ZnO particles was prepared; we composed quantum dots of good biocompatible by ultrasonic-assistant method; we marked the polymer by quantum dots, observe its distribution in the body of mice, analyzed it metabolization. The details are as follows:
1. Because of the catalysis of carbon nanotubes to H2O2 and the high iso-electric potential of ZnO nanopaticles, we prepared a glucosesensor based on the synergistic action of these two kinds of materials. By using nanomaterials, the performance of the sensor was remarkably improved. This sensor can provide an ideal microenvironment for enzyme and prevent the leakage of enzyme. The sensor can detect the glucose in solution with high sensitivity, good repeatability and anti-interference.
2. We prepared CdSe and CdSe/CdS quantum dots in aqueous solution using ultrasonic-assistant method. These quantum dots appeared good water-solubility and biocompatibility by linking mercaptoacetic acid. These quantum dots can be connected to biomolecule such as antibody, protein, DNA, which will show large potential application in immunofluorscence and biological molecular marking.
3. We connected CdSe/ZnS quantum dots to polymer molecular poly-PTX as fluorescence labels, analyzed the size, pattern of the marked polymer by particle sizing systems, TEM and so on. Then we injected the polymer solution to the body of mice, made the frozen section of the tissues, observed the distribution of the polymer by a high resolution fluorescence microscope. We analyzed the preliminary target of the poly-PTX and its metabolization in mice's body, the results showed that the quantum dots are perfect fluorescence labels of high efficiency, low toxicity and long-time stability. We made a basic exploration of the application of quantum dots in biological imaging.
1由于碳纳米管对过氧化氢的催化性以及氧化锌的高等电位,制备了基于此两种材料协同作用的葡萄糖传感器。纳米材料的应用,极大的提高了传感器的性能,该传感器为生物活性酶提供了一个良好的微环境并有效地防止酶泄漏,能检测溶液中葡萄糖的存在并具有较高的灵敏度、抗干扰性,及其较好的重复性。
2以超声辅助的方法,水相制备了CdSe和CdSe/CdS量子点,该量子点具有较好的水溶性,通过接枝巯基乙酸(MAA)使量子点具备较好的生物相容性,能够与常见生物分子如抗体,蛋白,DNA等相连,在免疫荧光分析,生物分子标记方面有很大的应用价值。
3将CdSe/ZnS量子点标记到载有药物的聚合物分子Poly-PTX上,用粒径仪,TEM等研究粒子的大小及形貌,将标记了量子点的聚合物注射到裸鼠体内,制作裸鼠各组织的冰冻切片,利用高分辨率荧光显微镜观察聚合物分子在裸鼠体内各个组织的分布,对Poly-PTX做一个初步的靶向分析,并分析其在裸鼠体内的代谢情况,实验结果表明量子点是一种高效、低毒、稳定的荧光标记物。本实验对量子点在生物成像领域的应用作了初步的探索。
When the size of some substances decrease to nanometre scale, they will show many special properties owing to their quantum confinement effect, surface and interface effect, small size effect and macroscopic quantum effect, which will be widely used in electronic science, material science, optics and biology. Recently, nanomaterials are more and more attractive because of their application in biosensor and biodetection. In this thesis, a biosensor based on synergistic action of carbon nanotubes and ZnO particles was prepared; we composed quantum dots of good biocompatible by ultrasonic-assistant method; we marked the polymer by quantum dots, observe its distribution in the body of mice, analyzed it metabolization. The details are as follows:
1. Because of the catalysis of carbon nanotubes to H2O2 and the high iso-electric potential of ZnO nanopaticles, we prepared a glucosesensor based on the synergistic action of these two kinds of materials. By using nanomaterials, the performance of the sensor was remarkably improved. This sensor can provide an ideal microenvironment for enzyme and prevent the leakage of enzyme. The sensor can detect the glucose in solution with high sensitivity, good repeatability and anti-interference.
2. We prepared CdSe and CdSe/CdS quantum dots in aqueous solution using ultrasonic-assistant method. These quantum dots appeared good water-solubility and biocompatibility by linking mercaptoacetic acid. These quantum dots can be connected to biomolecule such as antibody, protein, DNA, which will show large potential application in immunofluorscence and biological molecular marking.
3. We connected CdSe/ZnS quantum dots to polymer molecular poly-PTX as fluorescence labels, analyzed the size, pattern of the marked polymer by particle sizing systems, TEM and so on. Then we injected the polymer solution to the body of mice, made the frozen section of the tissues, observed the distribution of the polymer by a high resolution fluorescence microscope. We analyzed the preliminary target of the poly-PTX and its metabolization in mice's body, the results showed that the quantum dots are perfect fluorescence labels of high efficiency, low toxicity and long-time stability. We made a basic exploration of the application of quantum dots in biological imaging.
引文
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[2]Davis J, Vaughan D H, Cardosi M F. Enzyme Microb Tech,1995,17:1030-1035
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[4]Li J, Tan S N, Ge H. Anal. Chim. Acta,1996,335:137-145
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[7]张玲.中国科学技术大学博士学位论文(Doctoral Dissertation of The University of Science and Technology of China),2007
[8]麦智彬,谭学才,邹小勇.分析测试学报(Journal of Instrumental Analysis), 2006,25:120-125
[9]Yun Y H, Dong Z Y, Shanov V, et al. Nano Today,2007,2:30-37
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[15]段大雪,楚霞.化学传感器,2006,26：29-33
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[20]He P L, Hu N F. Electroanalysis [J],2004,16:1122
[21]He P L, Hu N F, Rusling J F. Langmuir [J],2004,20:722.
[22]Sapsford K E, Medintz I L, Golden I P, et al. Surface-Immobilized Self-Assembled Protein-Based Quantum Dot Nanoassemblies[J]. Langmuir,2004, 20(18):7720-7728.
[23]Dai Z, Kawde A N, Xiang Y, Belle J T L, Gerlach J, Bhavanandan V P, Joshi L, Wang J. J. Am. Chem. Soc.[J],2006,128:10018.
[24]Blanc Beguin F, Nabily S, Gieraltowski J, et al. Cytotoxicity and GMI bio-sensor detection of maghemite nanoparticles internalized into cells [J]. Journal of Magnetism and Magnetic Materials,2009,321(3):192-197.
[25]Zhang Y, Wang H, Yan B, et al. A reusable piezoelectric immunosensor using antibody-adsorbed magnetic nanocomposite[J]. J Immunol Methods,2008,332(1-2): 103-111.
[26]Guo S, Li J, Wang E. High-density gold nanoparticles supposed on a [Ru(bpy)3]2+-doped silica/Fe3O4 nanocomposite:facile prepa-ration, magnetically induced immobilization, and applications in ECL detection[J]. Chem Asian J,2008, 3(8-9):1544-1548.
[1]L. C. Clark, C. Lyons, Electrode systems for continuous monitoring in cardiovascular surgery, Ann. N.Y. Acad.Sci.102(1962)29-45
[2]J. B. Jia, B. Q. Wang, A. G Wu, G. G Cheng, Z. Li, S. J. Dong, A method to construct a third-generation horseradish peroxidase biosensor: self-assemblyng gold nanoparticles to three-demensional sol-gel network, Anal.Chem.74(2002) 2217-2223
[3]S. Zhang, N. Wang, Y. Niu, C. Sun, Immobilization of glucose oxidase on gold nanoparticles modified Au electrode for the construction of biosensor, Sens. Actuators B Chem.,109(2005) 367-374
[4]S. Zhao, K. Zhang, Y. Bai, W. W. Yang, C. Sun, Glucose oxidase/colloidal gold nanoparticles immobilized in Nafion film on glassy carbon electrode:Direct electron transfer and electrocatalysis Bioelectrochemistry,69(2006)158-163
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