量子点阵列图谱用于不同核酸碱基与血清蛋白的区分检测

发布时间：2018-03-28 18:04

本文选题：阵列图谱技术　切入点：量子点　出处：《湖南大学》2015年硕士论文

【摘要】：阵列图谱分析技术,又称为“化学鼻/舌头”,采用人工模拟嗅觉/味觉系统的传感模式,构建阵列图谱位点,基于不同传感位点的微分差异,实现目标物的区分与检测。该方法具有简单快速、高通量、便于微型组装等特点,在食品安全、生物医学检测、环境监测等领域具有潜在的应用前景。目前,纳米材料因为具有高比表面积、便于生物修饰与功能化等特点,将纳米材料应用于阵列图谱探针的构建,为阵列图谱技术的发展提供了新的契机。量子点(QDs)具有独特的尺寸依赖的光物理性质,作为优异的荧光探针,已广泛应用于化学生物传感、生物医学成像、化学催化等领域。本论文以阵列图谱分析作为出发点,采用量子点荧光纳米颗粒作为信号探针,发展对不同核酸碱基、血清蛋白质等重要生物分子的区分与检测技术,主要开展了以下两个方面的工作:1.基于不同核酸碱基对量子点激子能量转移效应的微分差异,发展了量子点的荧光阵列图谱方法,实现了不同核酸碱基的识别与区分。分别以巯基乙酸、2-二甲胺基乙硫醇盐酸盐、巯基乙胺和N-乙酰基-L-半胱氨酸四种不同配体合成了不同表面功能化的水溶性量子点,量子点分别与ATCGU五种核酸碱基构成阵列图谱。核酸碱基诱导量子点发生团聚与自组装,引起量子点之间的激子能量转移效应,导致量子点发生不同程度的荧光淬灭与发射波长红移。以量子点荧光强度为响应信号,基于不同量子点与不同核酸碱基的微分差异作用,采用线性判别分析,实现了五种不同核酸碱基的识别,并构建了核酸碱基区分的识别条形码;基于该方法进一步实现了不同稀有核酸碱基的快速区分。在该量子点阵列图谱中,量子点既作为荧光响应单元,又作为识别单元,提供了一种简单和无标记的生物传感方法,为量子点的生物传感和阵列图谱技术提供了新的平台。2.基于不同血清蛋白对于量子点纳米胶束的刺激响应作用,发展了量子点荧光共振能量转移的荧光阵列图谱方法,实现了不同血清蛋白质的识别与区分。4种不同表面活性剂(十二烷基磺酸钠、十二烷基肌氨酸钠、十六烷基三甲基溴化铵、十二烷基三甲基溴化铵)与4种聚电解质(聚甲基丙烯酸钠、聚苯乙烯磺酸钠-马来酸共聚物、聚二烯丙基二甲基氯化铵、聚烯丙基胺盐酸盐)进行组合,基于超分子自组装方法制备了7种纳米胶束,并同步包被量子点-Texas Red?DHPE的荧光共振能量供体受体对。5种不同的血清蛋白质(免疫球蛋白,人血清白蛋白,a-抗胰蛋白酶,纤维蛋白原,运铁蛋白)分别与7种纳米胶束作用,诱导纳米胶束发生组装变化,导致荧光染料的荧光变化。基于量子点纳米胶束与不同血清蛋白质的微分差异作用,构建了基于量子点的阵列图谱技术,结合线性判别分析,实现了5种血清蛋白质的阵列区分。对于未知浓度的蛋白质样品,采用吸光度浓度归一化方法,也达到了有效地辨别与区分。该阵列图谱方法设计灵活,信号灵敏,有望应用于血清蛋白质实际样品的检测与区分。基于量子点纳米胶束的可控组装,也为纳米胶束组装研究提供重要的参考价值。
[Abstract]:The array map analysis technology, also known as "chemical nose / tongue", the sensing mode of artificial olfactory and gustatory system, construction sites of different array atlas, differential sensing sites based on the implementation of differentiated and detection of the target. This method is simple and rapid, high throughput, easy to micro assembly, biomedical detection in food safety, and has potential applications in the field of environmental monitoring. At present, because the nanometer material has high specific surface area, easy modification and functional features of bio, nano materials used in construction of the map array probe, provide a new opportunity for the development of array technology. Quantum dots (QDs) with light depending on the size of the unique physical properties, excellent as a fluorescent probe, has been widely used in chemical and biological sensing, biomedical imaging, chemical catalysis and other fields. This paper analysis to array map As a starting point, by using quantum dot fluorescent nanoparticles as probe signal, development base on different nucleic acids, distinguish and detection of important biological molecules of serum proteins, mainly carried out the following two aspects: 1. based on different nucleic acid bases to differential difference transfer effect of excitons in quantum dots, fluorescence spectrum array development the method of quantum dots, can identify and distinguish different nucleobases. Respectively using thioglycolic acid, 2- two methylamino mercaptar hydrochloride, four different ligands Mercaptoethylamine and N- acetyl -L- cysteine was synthesized with different surface functionalization of water soluble quantum dots, quantum dots and ATCGU five arrays of nucleic acid bases Atlas of quantum dots aggregation and self-assembly induced by nucleic acid bases, caused by the exciton energy transfer between quantum effect, quantum dot fluorescence quenching leads to different degrees of destruction and Redshift of emission wavelength. The fluorescence intensity of quantum dots as response signal, differential difference of quantum dots with different nucleic acid bases based on the effect of using linear discriminant analysis, to achieve the recognition of five different nucleic acid bases, and the construction of the nucleic acid bases distinguish identification bar code; based on this method and further realize the rapid distinction between different nucleobases rare in the quantum dot arrays of quantum dots as fluorescent response units, and as recognition unit, provides a simple and label free biosensing method, fingerprint technology provides a new platform for.2. stimulation of different serum protein quantum dot nano micelles response based on quantum dots and biological sensor array the development of the column, fluorescent quantum dots array method of fluorescence resonance energy transfer, to achieve the recognition of different serum proteins and distinguish between different.4 surfactants (ten Two sodium dodecyl sulfate, twelve alkyl sarcosinate, sixteen alkyl three methyl bromide, twelve alkyl three methyl bromide) and 4 kinds of polyelectrolyte (poly methacrylic acid sodium polystyrene sulfonate - maleic acid copolymer, poly diallyl dimethyl ammonium chloride two, polyallylamine hydrochloride) combination the supramolecular self-assembly method of 7 kinds of nano micelles were prepared based on the synchronous and coated -Texas quantum dots Red? DHPE fluorescence resonance energy donor receptor on.5 in serum of different protein (immunoglobulin, human serum albumin, a- antitrypsin, fibrinogen and transferrin) respectively with 7 nano micelles effect induced by nano micelle assembly changes lead to fluorescence change of fluorescent dye. The differential difference of quantum dot nano micelles with different serum proteins based on constructed array mapping technique based on quantum dots, combined with linear discriminant Analysis of 5 serum protein array achieved distinction. For an unknown concentration of protein samples, the absorbance concentration normalization method, also can effectively identify and distinguish. The array mapping method of flexible design, signal sensitivity, detection and differentiation of serum protein is expected to be applied to the actual sample assembly. Controllable quantum dot nano micelles based on, but also provide an important reference value for the assembly of nano micelles.

【学位授予单位】：湖南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;O657.3

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