荧光纳米探针的构筑及生化分析应用
发布时间：2018-09-05 10:39
【摘要】：近来,荧光纳米探针的研究已受到全世界学者的普遍关注,并己被广泛被用于生化分析研究,如荧光成像,生物分子检测,环境中有毒有害物质的检测等,此类方法快速、简便、灵敏度高。本论文采用两种不同的方法构筑了多功能荧光纳米探针,将其分别用于基因输运的荧光示踪及环境中硝基苯类爆炸物的荧光分析检测。主要研究成果如下：1.癌症的基因治疗具有巨大的应用前景,但其负载、安全输运尤其是输运的实时示踪仍是研究的热点与难点。本工作采用自由基聚合方法合成了两亲正电高分子(聚-苯乙烯-甲基丙烯酸-(氯化1-烯丙基-3-甲基咪唑),P-St-MAA-AMIM),以该正电高分子为载体,以聚乙二醇一聚(乳酸-乙醇酸)(MPEG-PLGA)为辅助高分子,以荧光量子点ZnS:Mn2+为发光中心,通过简单的超声乳化,构筑了一种多功能纳米胶囊。既可用于基因的负载与输运,还可利用其荧光实现基因输运的实时荧光示踪与疗效评价。该功能纳米球尺寸小且均一,荧光稳定性好,生物相容性与水溶性好,对基因序列具有较高的负载效率,负载后的基因活性强,转染效率高。该方法发展了一种多功能基因纳米载体的制备新方法,并成功用于基因的负载、输运及荧光示踪。2.硝基苯类爆炸物的快速、灵敏、选择性检测是环境监测与公共安全领域普遍关注的热点,发展相应的灵敏检测方法至关重要。通过光诱导疏水性无机量子点原位聚合,一步得到了亲水性无机一有机杂化荧光纳米复合球,荧光纳米球的表面具有丰富氨基,亲水性好。由于三硝基苯酚TNP的UV-vis吸收光谱范围在300-450 nm,这对复合纳米球(NCs-NH2)的激发峰342 nm有显著的吸收,从而导致NCs-NH2纳米球591 nm处的荧光产生显著淬灭,可用于TNP的荧光定量检测,线性范围0.05- 8.0μg/mL,检测限为3.4 nM。而对于TNT,在pH11时,对纳米球的荧光无影响,pH=12或更高时,基于TNT与纳米球表面氨基间的电子转移效应及TNT负离子宽的UV-vis吸收(350-600 nm)共同作用,导致极低浓度的TNT也会对NCs-NH2纳米球591 nm处的荧光发生淬灭效应。且荧光强度与TNT浓度呈线性变换,可用于极低浓度TNT的定量分析。线性范围0.01-0.5μg/mL,检测限为2.2 nM。该方法发展了一种光诱导无机纳米晶聚合制备无机-有机复合纳米荧光探针的新方法,在不同pH条件下,实现了对TNP与TNT的同时选择性荧光分析。
[Abstract]:Recently, the research of fluorescent nanoprobes has been widely concerned by scholars all over the world, and has been widely used in biochemical analysis, such as fluorescence imaging, biomolecular detection, detection of toxic and harmful substances in the environment, and so on. It is simple and sensitive. In this paper, two different methods were used to construct multifunctional fluorescent nanoprobes, which were used for the fluorescence tracer of gene transport and the detection of nitrobenzene explosives in the environment. The main research results are as follows: 1. The gene therapy of cancer has great application prospect, but its load, safe transportation, especially the real-time tracer of transportation is still a hot and difficult point. A positive amphiphilic polymer (polystyrene-methacrylic acid) (1-allyl-3-methylimidazolium chloride) (P-St-MAA-MIM) was synthesized by free radical polymerization. A multifunctional nano-capsule was prepared by simple phacoemulsification with poly (ethylene glycol) -poly (lactic acid-glycolic acid) (MPEG-PLGA) as auxiliary polymer and fluorescent quantum dot (ZnS:Mn2) as luminescent center. It can be used not only for gene loading and transportation, but also for real-time fluorescence tracer and therapeutic evaluation of gene transport. The functional nanospheres are of small size, uniform size, good fluorescence stability, good biocompatibility and water solubility, high loading efficiency for gene sequence, strong gene activity after loading and high transfection efficiency. This method has developed a new method for the preparation of multifunctional gene nanoparticles, and has been successfully used in gene loading, transport and fluorescence tracer. 2. The rapid, sensitive and selective detection of nitrobenzene explosives is a hot topic in the field of environmental monitoring and public safety. In situ polymerization of hydrophobic inorganic quantum dots, the hydrophilic inorganic / organic hybrid fluorescent nanocomposite spheres were obtained in one step. The surface of the fluorescent nanospheres was rich in amino groups and had good hydrophilicity. Because the UV-vis absorption spectrum of trinitrophenol TNP is 300-450 nm, the excitation peak 342 nm of composite nanospheres (NCs-NH2) is significantly absorbed, which leads to the fluorescence quenching of NCs-NH2 nanospheres 591 nm, which can be used for the quantitative detection of TNP. The linear range is 0.05-8.0 渭 g / mL and the detection limit is 3.4 nM.. However, for TNT, at pH11, the fluorescence of nanospheres had no effect on the fluorescence of nanospheres at pH = 12 or higher, based on the electron transfer effect between TNT and amino groups on the surface of the nanospheres and the interaction of UV-vis absorption (350-600 nm) with TNT anion width. The fluorescence of NCs-NH2 nanospheres at 591 nm can also be quenched by very low concentration of TNT. The fluorescence intensity is linear with the concentration of TNT, which can be used for quantitative analysis of very low concentration TNT. The linear range is 0.01-0.5 渭 g / mL and the detection limit is 2.2 nM.. This method developed a new method of photoinduced inorganic nanocrystalline polymerization to prepare inorganic / organic composite nano-fluorescent probe. The simultaneous selective fluorescence analysis of TNP and TNT was realized under different pH conditions.
【学位授予单位】：北京化工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：O657.3;TB383.1
本文编号：2224023
[Abstract]:Recently, the research of fluorescent nanoprobes has been widely concerned by scholars all over the world, and has been widely used in biochemical analysis, such as fluorescence imaging, biomolecular detection, detection of toxic and harmful substances in the environment, and so on. It is simple and sensitive. In this paper, two different methods were used to construct multifunctional fluorescent nanoprobes, which were used for the fluorescence tracer of gene transport and the detection of nitrobenzene explosives in the environment. The main research results are as follows: 1. The gene therapy of cancer has great application prospect, but its load, safe transportation, especially the real-time tracer of transportation is still a hot and difficult point. A positive amphiphilic polymer (polystyrene-methacrylic acid) (1-allyl-3-methylimidazolium chloride) (P-St-MAA-MIM) was synthesized by free radical polymerization. A multifunctional nano-capsule was prepared by simple phacoemulsification with poly (ethylene glycol) -poly (lactic acid-glycolic acid) (MPEG-PLGA) as auxiliary polymer and fluorescent quantum dot (ZnS:Mn2) as luminescent center. It can be used not only for gene loading and transportation, but also for real-time fluorescence tracer and therapeutic evaluation of gene transport. The functional nanospheres are of small size, uniform size, good fluorescence stability, good biocompatibility and water solubility, high loading efficiency for gene sequence, strong gene activity after loading and high transfection efficiency. This method has developed a new method for the preparation of multifunctional gene nanoparticles, and has been successfully used in gene loading, transport and fluorescence tracer. 2. The rapid, sensitive and selective detection of nitrobenzene explosives is a hot topic in the field of environmental monitoring and public safety. In situ polymerization of hydrophobic inorganic quantum dots, the hydrophilic inorganic / organic hybrid fluorescent nanocomposite spheres were obtained in one step. The surface of the fluorescent nanospheres was rich in amino groups and had good hydrophilicity. Because the UV-vis absorption spectrum of trinitrophenol TNP is 300-450 nm, the excitation peak 342 nm of composite nanospheres (NCs-NH2) is significantly absorbed, which leads to the fluorescence quenching of NCs-NH2 nanospheres 591 nm, which can be used for the quantitative detection of TNP. The linear range is 0.05-8.0 渭 g / mL and the detection limit is 3.4 nM.. However, for TNT, at pH11, the fluorescence of nanospheres had no effect on the fluorescence of nanospheres at pH = 12 or higher, based on the electron transfer effect between TNT and amino groups on the surface of the nanospheres and the interaction of UV-vis absorption (350-600 nm) with TNT anion width. The fluorescence of NCs-NH2 nanospheres at 591 nm can also be quenched by very low concentration of TNT. The fluorescence intensity is linear with the concentration of TNT, which can be used for quantitative analysis of very low concentration TNT. The linear range is 0.01-0.5 渭 g / mL and the detection limit is 2.2 nM.. This method developed a new method of photoinduced inorganic nanocrystalline polymerization to prepare inorganic / organic composite nano-fluorescent probe. The simultaneous selective fluorescence analysis of TNP and TNT was realized under different pH conditions.
【学位授予单位】：北京化工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：O657.3;TB383.1
【共引文献】
相关期刊论文 前1条
1 贺冬秀;王丹霞;全文捷;喻翠云;;功能化量子点在生物分析,生物成像和靶向运输中的应用（英文）[J];中南医学科学杂志;2015年05期
相关硕士学位论文 前2条
1 葛艳秋;基于DNA功能化纳米信标的电化学免疫传感方法研究[D];大连理工大学;2014年
2 袁玲;与氧化应激相关的生物分子的电化学传感[D];南京师范大学;2014年
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