發(fā)布時(shí)間：2018-03-11 16:49

  本文選題：金屬納米微粒　切入點(diǎn)：局域表面等離子共振　出處：《南京郵電大學(xué)》2016年博士論文　論文類型：學(xué)位論文

【摘要】：納米生物材料作為納米科技領(lǐng)域中的一個(gè)主要方面,其技術(shù)的進(jìn)步和發(fā)展引領(lǐng)著生物醫(yī)學(xué)、藥物運(yùn)輸?shù)阮I(lǐng)域的蓬勃發(fā)展。利用生物分子對(duì)納米材料進(jìn)行修飾,可以結(jié)合生物分子的識(shí)別能力與納米材料的優(yōu)良特性,構(gòu)建出多種新型的功能性納米材料探針,在生物傳感、靶向治療以及光電材料等方面具有重要的意義。納米生物材料領(lǐng)域的關(guān)鍵問(wèn)題之一是如何構(gòu)建功能性納米材料探針?lè)肿右猿浞掷蒙锓肿?如:DNA、RNA以及蛋白質(zhì)等)和納米材料結(jié)合的優(yōu)勢(shì)。本論文旨在基于金屬納米顆粒構(gòu)建探針?lè)肿佑糜诤怂岱肿拥臋z測(cè)。論文的研究?jī)?nèi)容包括以下四部分:1.單顆粒銀納米立方體探針的制備及其對(duì)早期肺癌標(biāo)志物miRNA的實(shí)時(shí)檢測(cè)MicroRNA作為腫瘤標(biāo)志物,調(diào)控著很多細(xì)胞內(nèi)過(guò)程。然而,由于其序列短、含量低等缺陷,很難開(kāi)發(fā)簡(jiǎn)單、快速、廉價(jià)的檢測(cè)方法。我們?cè)O(shè)計(jì)了巰基單鏈DNA (ssDNA)修飾的單顆粒銀納米立方體(AgNC)探針?lè)肿?并基于此探針?lè)肿訕?gòu)建了等離子體納米生物傳感器用于microRNA-21 (miR-21)的檢測(cè)。我們首次通過(guò)原位實(shí)驗(yàn)成功地實(shí)時(shí)觀測(cè)到核酸分子在單個(gè)AgNC表面雜交的過(guò)程。實(shí)驗(yàn)結(jié)果及時(shí)域有限差分方法(FDTD)仿真結(jié)果都表明了 AgNCs局域表面等離子體共振(LSPR)峰的紅移是由于ssDNA分子與靶分子miR-21雜交引起AgNCs表面折射率(RI)增大的結(jié)果。隨著靶分子miR-21濃度的增大,AgNCs的LSPR峰紅移量增加,對(duì)靶分子miR-21的檢測(cè)靈敏度可達(dá)到1fM。此外,通過(guò)FDTD仿真模型,成功的解釋了反應(yīng)的機(jī)理,每一個(gè)AgNC都可以作為一個(gè)納米級(jí)傳感探針用于早期肺癌標(biāo)志物miR-21的檢測(cè)。2.單顆粒金銀核殼納米立方體探針的制備及其對(duì)miRNA單分子水平的檢測(cè)和邏輯門(mén)的構(gòu)建在許多生物系統(tǒng)中,單分子水平的研究可以揭示分子間的相互作用、動(dòng)力學(xué)以及構(gòu)象的細(xì)微變化。目前用于單分子水平檢測(cè)的可行方法通常需要熒光分子作標(biāo)記,然而熒光探針容易發(fā)生光漂白等現(xiàn)象且信噪比較低。因此,我們?cè)O(shè)計(jì)了四面體結(jié)構(gòu)的DNA (tsDNA)分子修飾的單顆粒金銀核殼納米立方體(Au@AgNC)探針?lè)肿?并基于此探針?lè)肿訕?gòu)建了“智能”等離子體納米生物傳感器用于單分子水平microRNA-21 (miR-21)以及核酸內(nèi)切酶(KpnI和StuI)的檢測(cè)。單個(gè)miR-21與tsDNA的雜交過(guò)程可引起Au@Ag NC的局域表面等離子體共振(LSPR)峰發(fā)生約0.4nm的紅移。這種“智能”等離子體納米生物傳感器不僅可以實(shí)現(xiàn)對(duì)miR-21的超靈敏檢測(cè),檢測(cè)動(dòng)態(tài)范圍從1 aM到1 nM,而且成功地執(zhí)行了以miR-21、KpnI和StuI響應(yīng)為模型的基于tsDNA的邏輯運(yùn)算以及生物存儲(chǔ)器。3.基于單層過(guò)渡金屬二硫化物納米片探針的制備及其對(duì)DNA的雙元檢測(cè)快速、靈敏的同時(shí)實(shí)現(xiàn)對(duì)多種靶分子的檢測(cè)是基因表達(dá)和分子診斷領(lǐng)域的挑戰(zhàn)性問(wèn)題之一。針對(duì)這一問(wèn)題,我們?cè)O(shè)計(jì)了基于單層過(guò)渡金屬二硫化物(TMD,包括MoS2、TiS2和TaS2)納米片的探針?lè)肿佑糜贒NA分子的檢測(cè)。利用TMD納米片的高熒光猝滅能力以及對(duì)于染料分子標(biāo)記的單鏈DNA (ssDNA)和雙鏈DNA (dsDNA)不同的親和力的特性構(gòu)建的新型熒光傳感平臺(tái)。通過(guò)比較發(fā)現(xiàn)基于單層TaS2納米片的DNA傳感器對(duì)染料分子標(biāo)記的ssDNA具有最高的猝滅效率(99%)和最低的檢測(cè)限(0.05 nM)。重要的是,利用基于單層TaS2納米片的DNA傳感器首次實(shí)現(xiàn)了同時(shí)對(duì)靶分子H1N1和H5N1的檢測(cè)。4.基于DNA為合成模板的銀納米簇探針的制備及其對(duì)DNA的雙元檢測(cè)與有機(jī)染料相比,金屬納米顆粒具有穩(wěn)定性強(qiáng)、熒光強(qiáng)度高以及熒光壽命長(zhǎng)等優(yōu)點(diǎn)。我們進(jìn)一步地設(shè)計(jì)了一種基于DNA為模板合成的熒光銀納米簇(AgNCs)的新型免標(biāo)記的分子信標(biāo)(MB)探針,用于DNA的雙元檢測(cè)。以兩條不同的DNA序列為模板合成的發(fā)射光譜峰分別位于507 nm的綠光Ag NCs和597 nm的橙光Ag NCs作為熒光信號(hào),可以實(shí)現(xiàn)廉價(jià)、免標(biāo)記的對(duì)靶分子H1N1和H5N1的同時(shí)檢測(cè),檢測(cè)限為25nM。
[Abstract]:Nanometer biological materials as one of the main aspects in the field of nanotechnology, the progress and development of the technology leads the vigorous development of biomedicine, drug transport and other fields. The use of biological molecular modification of nano materials, excellent properties can be combined with the ability to identify biological molecules and nano materials, build a variety of new functional nanomaterials probe in the biosensing, targeted therapy and photoelectric materials has important significance. One of the key issues in the field of nano biomaterials is how to construct the molecular probe of functional nanomaterials to make full use of biological molecules (such as: DNA, RNA and proteins) and combined with nano materials. The advantages of this thesis metal nano particles construction of molecular probe for the detection of nucleic acid molecules. Based on the research contents of this paper include the following four parts: 1. single particle silver nanocubes probe system Preparation and real-time detection of MicroRNA in early lung cancer marker miRNA as a tumor marker, regulates many cellular processes. However, due to its short sequences, low content of defects, it is difficult to develop a simple, fast and cheap detection method. We design the mercapto single stranded DNA (ssDNA) single particle (modified silver nanocubes AgNC) and the probe molecule probe molecule is constructed based on plasma nano biosensor for detection of microRNA-21 (miR-21). For the first time through the process of in situ experiments we successfully observed real-time nucleic acid molecules in a single AgNC surface hybridization. The experimental results and the finite difference time domain method (FDTD) simulation results show that the localized surface plasmon AgNCs resonance (LSPR) peak red shift is due to the ssDNA molecule and the target molecule miR-21 hybridization induced AgNCs surface index (RI) increased. With the increase of the concentration of target molecules of miR-21 LSPR, peak redshift AgNCs increases, the detection sensitivity of the target molecules of miR-21 can reach 1fM. in addition, through the FDTD simulation model successfully explains the reaction mechanism, each AgNC can be used as a nano sensing probe for early detection of lung cancer marker.2. miR-21 single particle silver core-shell nano cube the preparation of miRNA probe and single molecule detection and logic gate construction in many biological systems, studies on single molecular level reveals the interaction between molecules, dynamics and conformational changes. At present for the feasible method of single molecule level detection usually requires a fluorescent molecular marking, but prone to fluorescent probe photobleaching phenomenon and the SNR is low. Therefore, we design the tetrahedral structure of DNA (tsDNA) single particle silver core-shell nano cube molecular modification (Au@AgNC) probe This molecule, and molecular probe was constructed based on the "smart" plasma nanobiosensors for single molecule level microRNA-21 (miR-21) and endonucleases (KpnI and StuI) in the detection of hybridization. The single miR-21 and tsDNA Au@Ag NC can cause local surface plasmon resonance (LSPR) peak has a red shift of about 0.4nm. This "smart" plasma nanobiosensors can not only realize the ultra sensitive detection of miR-21, detection of dynamic range from 1 aM to 1 nM, and the successful implementation of miR-21, KpnI and StuI in response to the model based on tsDNA logic and biological memory.3. monolayer transition metal sulfide two probes were prepared based on nano film and the dual DNA assay is rapid, sensitive and realize the detection of a variety of target molecules is one of the most challenging problems in the field of molecular diagnostics and gene expression. In view of this question Questions, we designed a single transition metal sulfide based on two (TMD, including MoS2, TiS2 and TaS2) molecular probe nanosheets for the detection of DNA molecules. The high fluorescence quenching ability by TMD nanosheets and dye molecular markers for single stranded DNA (ssDNA) and double stranded DNA (dsDNA) a new fluorescent sensing platform different affinity characteristics. By comparing the ssDNA DNA sensor single-layer TaS2 nanosheets of dye molecular markers based on the highest quenching efficiency (99%) and the lowest detection limit (0.05 nM). It is important that the use of DNA sensor based on single TaS2 nanosheets first realized the simultaneous detection of.4. of target molecules H1N1 and H5N1 silver nanoclusters probe synthesis template preparation and dual DNA detection compared with organic dyes for DNA based metal nanoparticles has strong stability, high fluorescence intensity and long fluorescence lifetime etc. We have further advantages. Based on the design of a DNA as a template for the synthesis of fluorescent silver nanoclusters (AgNCs) of the new label free molecular beacon (MB) dual probe for the detection of DNA. With two different DNA sequences as the template synthesis and emission peaks are located at 507 nm green light Ag NCs and 597 nm orange light Ag NCs as fluorescence signal, can realize the detection of cheap, label free target molecules of H1N1 and H5N1 at the same time, the detection limit is 25nM.

【學(xué)位授予單位】：南京郵電大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R734.2;TB383.1

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相關(guān)期刊論文 前2條

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本文編號(hào)：1599041