ZnO/Au纳米复合物光学性质及利用其拉曼信号实施对蛋白分子的检测研究
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摘要
氧化锌(ZnO)是一种具有六方结构的Ⅱ-Ⅵ族宽带隙半导体材料,室温下带隙宽度高达3.3eV。由于氧化锌具有较高的激子束缚能(60meV),保证了其在室温下较强的激子发光,因而被认为是制作紫外半导体激光器的合适材料。但是其材料本身也具有一些缺陷,金属半导体材料能够利用其金属特性弥补半导体材料的缺陷,使其获得优良的特性,这为材料进一步在器件上应用提供了材料基础。ZnO除了做器件材料外,在生物医药、涂料、食品、化妆品等与人们生活密切相关的各个领域被广泛关注。
本文通过对ZnO及ZnO/Au纳米复合物的制备和光学性质研究,实现了ZnO/Au纳米复合物对未知蛋白分子的超灵敏检测。具体研究内容如下:
1.还原法制备水溶性ZnO/Au纳米复合粒子,并研究其光学性质。透射电镜表征了ZnO/Au纳米粒子的晶体形貌和结构;紫外吸收光谱表明ZnO/Au纳米复合物的形成,在250-400 nm波长范围内,对应ZnO的激子吸收,在400-700 nm波长范围内,对应Au的等离子体吸收。结合光谱分析和结构分析确定ZnO和Au之间存在着界面电荷转移。光致发光谱显示了Au对ZnO发光性质的影响。在325nm激光激发下观察到了ZnO/Au纳米复合物的共振拉曼光谱随Au含量变化的规律。
2.ZnO/Au纳米复合物的生物相容性处理研究,通过改变Au与ZnO的复合的比例关系确定能够使ZnO在水中稳定性好的ZnO,Au的最优比例关系。
3.采用三明治夹心结构的免疫层析原理研究利用ZnO的共振拉曼信号作为指纹特征来检测未知生物分子。ZnO/Au纳米复合物与人IgG偶联后可以作为一种探针,具有较强的共振拉曼信号。这种方法有较高的检测灵敏度.
4.通过改变溶液的浓度及盐离子参数获得不同形貌的ZnO/Au纳米复合物,并研究其光学结构特性。
Zinc oxide is aⅡ-Ⅵwide band-gap(3.3eV) semiconductor with wurtzite crystal structure.Due to the large exciton binding energy of 60meV,it is regarded as one of the most promising materials for fabricating efficient ultraviolet(UV) and blue light emitting devices.But the materials have some defects themselves, metal-semiconducting material can remedy defects of semiconducting material by the properties of metals.They would get excellent properties and provide base of the future application in devices.Otherwise,ZnO has also been widely focused in many fields such as medicine,solar energy cell,food,cosmetics and so on.
In this thesis,ZnO nanomaterials were chosen as the subject of investigation, which was focused on the synthesis and characterization of water-soluble ZnO/Au nanocomposites.ZnO/Au nanocomposites were used for detecting unknown protein molecule base on Resonant Raman Scattering.The details are as follows:
(1) The ZnO/Au nanocomposites were successful prepared by reduction reaction. The size distribution and morphology of the ZnO/Au nanocomposites were characterized by the TEM.The UV-vis absorption spectra of ZnO/Au nanocomposites exhibited well-defined exciton band of ZnO quantum dots (QDs) in the ultraviolet region(250-400 nm).The plasmon absorption band of Au nanocrystals was observed in the visible region(400-700 nm).Combined with optical spectra and structure analysis,the electron transfer between ZnO and Au may be thought.The photoluminescence of ZnO/Au nanocomposites showed the effect of Au on ZnO nanocomposites.Furthermore,the resonance Raman scattering of the ZnO/Au nanocomposites was observed with excitation of 325 nm light from He—Cd laser.The amount of Au coating on the surface of ZnO is important for obtaining different intensity of enhanced Raman scattering.
(2) The biocompatible ZnO/Au nanocomposites were studied by changing the the amount of Au on ZnO nanoparticles.Au layer on ZnO surface has great effect on the ZnO in aqueous solution due to the interaction between ligand on Au and water molecule.
(3) A probe for the target protein was constructed by binding the ZnO/Au nanoparticles to secondary protein by eletrostatic interaction.The detection of proteins was achieved by an antibody-based sandwich assay.A first antibody,which could be specifically recognized by target protein,was attached to a solid silicon surface.The ZnO/Au protein probe could specifically recognize and bind to the complex of the target protein and first antibody.This method on the resonant Raman scattering signal of ZnO nanoparticles showed good selectivity and sensitivity for the target protein.
(4) Different morphology of ZnO/Au nanocomposites was obtained by changing the salt concentration of solution.We also study on the optical properties of these nanocomposites.
本文通过对ZnO及ZnO/Au纳米复合物的制备和光学性质研究,实现了ZnO/Au纳米复合物对未知蛋白分子的超灵敏检测。具体研究内容如下:
1.还原法制备水溶性ZnO/Au纳米复合粒子,并研究其光学性质。透射电镜表征了ZnO/Au纳米粒子的晶体形貌和结构;紫外吸收光谱表明ZnO/Au纳米复合物的形成,在250-400 nm波长范围内,对应ZnO的激子吸收,在400-700 nm波长范围内,对应Au的等离子体吸收。结合光谱分析和结构分析确定ZnO和Au之间存在着界面电荷转移。光致发光谱显示了Au对ZnO发光性质的影响。在325nm激光激发下观察到了ZnO/Au纳米复合物的共振拉曼光谱随Au含量变化的规律。
2.ZnO/Au纳米复合物的生物相容性处理研究,通过改变Au与ZnO的复合的比例关系确定能够使ZnO在水中稳定性好的ZnO,Au的最优比例关系。
3.采用三明治夹心结构的免疫层析原理研究利用ZnO的共振拉曼信号作为指纹特征来检测未知生物分子。ZnO/Au纳米复合物与人IgG偶联后可以作为一种探针,具有较强的共振拉曼信号。这种方法有较高的检测灵敏度.
4.通过改变溶液的浓度及盐离子参数获得不同形貌的ZnO/Au纳米复合物,并研究其光学结构特性。
Zinc oxide is aⅡ-Ⅵwide band-gap(3.3eV) semiconductor with wurtzite crystal structure.Due to the large exciton binding energy of 60meV,it is regarded as one of the most promising materials for fabricating efficient ultraviolet(UV) and blue light emitting devices.But the materials have some defects themselves, metal-semiconducting material can remedy defects of semiconducting material by the properties of metals.They would get excellent properties and provide base of the future application in devices.Otherwise,ZnO has also been widely focused in many fields such as medicine,solar energy cell,food,cosmetics and so on.
In this thesis,ZnO nanomaterials were chosen as the subject of investigation, which was focused on the synthesis and characterization of water-soluble ZnO/Au nanocomposites.ZnO/Au nanocomposites were used for detecting unknown protein molecule base on Resonant Raman Scattering.The details are as follows:
(1) The ZnO/Au nanocomposites were successful prepared by reduction reaction. The size distribution and morphology of the ZnO/Au nanocomposites were characterized by the TEM.The UV-vis absorption spectra of ZnO/Au nanocomposites exhibited well-defined exciton band of ZnO quantum dots (QDs) in the ultraviolet region(250-400 nm).The plasmon absorption band of Au nanocrystals was observed in the visible region(400-700 nm).Combined with optical spectra and structure analysis,the electron transfer between ZnO and Au may be thought.The photoluminescence of ZnO/Au nanocomposites showed the effect of Au on ZnO nanocomposites.Furthermore,the resonance Raman scattering of the ZnO/Au nanocomposites was observed with excitation of 325 nm light from He—Cd laser.The amount of Au coating on the surface of ZnO is important for obtaining different intensity of enhanced Raman scattering.
(2) The biocompatible ZnO/Au nanocomposites were studied by changing the the amount of Au on ZnO nanoparticles.Au layer on ZnO surface has great effect on the ZnO in aqueous solution due to the interaction between ligand on Au and water molecule.
(3) A probe for the target protein was constructed by binding the ZnO/Au nanoparticles to secondary protein by eletrostatic interaction.The detection of proteins was achieved by an antibody-based sandwich assay.A first antibody,which could be specifically recognized by target protein,was attached to a solid silicon surface.The ZnO/Au protein probe could specifically recognize and bind to the complex of the target protein and first antibody.This method on the resonant Raman scattering signal of ZnO nanoparticles showed good selectivity and sensitivity for the target protein.
(4) Different morphology of ZnO/Au nanocomposites was obtained by changing the salt concentration of solution.We also study on the optical properties of these nanocomposites.
引文
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[3]张阳德.纳米生物分析化学与分子生物学[M].北京:化学工业出版社,2005.7.
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[18]Dou X,Takama T,Yamaguchi Y,et al.Enzyme Immunoassay Utilizing Surface-Enhanced Raman Scattering of the Enzyme Reaction Product[J].Anal Chem,1997,69:1492.
[19]徐抒平,王连英,徐蔚青.SERS标记纳米粒子用于免疫识别[J].高等学校化学学报,2003,24(5):900.
[20]Manfait M,Morjani h,Millot J M,et al.Drug-target interactions on a single living cell:an approach by optical microspectroscopy[J].Proc SPIE-lnt Soc Opt Eng.,1990,1403:695-709.
[21]Manfait M,Morjani H,Nabiev I.Molecular events on simple living cancer cells as studied by spectrofluorometry and micro-SERS Raman spectroscopy [J].J Cell Pharmacol 1992,3:120-125.
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