熒光納米探針的構(gòu)筑及生化分析應(yīng)用
發(fā)布時間：2018-09-05 10:39
【摘要】：近來,熒光納米探針的研究已受到全世界學(xué)者的普遍關(guān)注,并己被廣泛被用于生化分析研究,如熒光成像,生物分子檢測,環(huán)境中有毒有害物質(zhì)的檢測等,此類方法快速、簡便、靈敏度高。本論文采用兩種不同的方法構(gòu)筑了多功能熒光納米探針,將其分別用于基因輸運(yùn)的熒光示蹤及環(huán)境中硝基苯類爆炸物的熒光分析檢測。主要研究成果如下：1.癌癥的基因治療具有巨大的應(yīng)用前景,但其負(fù)載、安全輸運(yùn)尤其是輸運(yùn)的實(shí)時示蹤仍是研究的熱點(diǎn)與難點(diǎn)。本工作采用自由基聚合方法合成了兩親正電高分子(聚-苯乙烯-甲基丙烯酸-(氯化1-烯丙基-3-甲基咪唑),P-St-MAA-AMIM),以該正電高分子為載體,以聚乙二醇一聚(乳酸-乙醇酸)(MPEG-PLGA)為輔助高分子,以熒光量子點(diǎn)ZnS:Mn2+為發(fā)光中心,通過簡單的超聲乳化,構(gòu)筑了一種多功能納米膠囊。既可用于基因的負(fù)載與輸運(yùn),還可利用其熒光實(shí)現(xiàn)基因輸運(yùn)的實(shí)時熒光示蹤與療效評價。該功能納米球尺寸小且均一,熒光穩(wěn)定性好,生物相容性與水溶性好,對基因序列具有較高的負(fù)載效率,負(fù)載后的基因活性強(qiáng),轉(zhuǎn)染效率高。該方法發(fā)展了一種多功能基因納米載體的制備新方法,并成功用于基因的負(fù)載、輸運(yùn)及熒光示蹤。2.硝基苯類爆炸物的快速、靈敏、選擇性檢測是環(huán)境監(jiān)測與公共安全領(lǐng)域普遍關(guān)注的熱點(diǎn),發(fā)展相應(yīng)的靈敏檢測方法至關(guān)重要。通過光誘導(dǎo)疏水性無機(jī)量子點(diǎn)原位聚合,一步得到了親水性無機(jī)一有機(jī)雜化熒光納米復(fù)合球,熒光納米球的表面具有豐富氨基,親水性好。由于三硝基苯酚TNP的UV-vis吸收光譜范圍在300-450 nm,這對復(fù)合納米球(NCs-NH2)的激發(fā)峰342 nm有顯著的吸收,從而導(dǎo)致NCs-NH2納米球591 nm處的熒光產(chǎn)生顯著淬滅,可用于TNP的熒光定量檢測,線性范圍0.05- 8.0μg/mL,檢測限為3.4 nM。而對于TNT,在pH11時,對納米球的熒光無影響,pH=12或更高時,基于TNT與納米球表面氨基間的電子轉(zhuǎn)移效應(yīng)及TNT負(fù)離子寬的UV-vis吸收(350-600 nm)共同作用,導(dǎo)致極低濃度的TNT也會對NCs-NH2納米球591 nm處的熒光發(fā)生淬滅效應(yīng)。且熒光強(qiáng)度與TNT濃度呈線性變換,可用于極低濃度TNT的定量分析。線性范圍0.01-0.5μg/mL,檢測限為2.2 nM。該方法發(fā)展了一種光誘導(dǎo)無機(jī)納米晶聚合制備無機(jī)-有機(jī)復(fù)合納米熒光探針的新方法,在不同pH條件下,實(shí)現(xiàn)了對TNP與TNT的同時選擇性熒光分析。
[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.
【學(xué)位授予單位】：北京化工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：O657.3;TB383.1
本文編號：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.
【學(xué)位授予單位】：北京化工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：O657.3;TB383.1
【共引文獻(xiàn)】
相關(guān)期刊論文 前1條
1 賀冬秀;王丹霞;全文捷;喻翠云;;功能化量子點(diǎn)在生物分析,生物成像和靶向運(yùn)輸中的應(yīng)用（英文）[J];中南醫(yī)學(xué)科學(xué)雜志;2015年05期
相關(guān)碩士學(xué)位論文 前2條
1 葛艷秋;基于DNA功能化納米信標(biāo)的電化學(xué)免疫傳感方法研究[D];大連理工大學(xué);2014年
2 袁玲;與氧化應(yīng)激相關(guān)的生物分子的電化學(xué)傳感[D];南京師范大學(xué);2014年
，本文編號：2224023
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