【作者】 许定花；

【导师】 金征宇； 胥传来；

【作者基本信息】 江南大学 ， 制糖工程， 2009， 硕士

【摘要】 由于量子点及碳纳米管(CNTs)独特的纳米特性及生物兼容性,它们在免疫分析领域被广泛应用。本论文分别构建了两种基于免疫反应的高灵敏检测方法。基于聚合量子点的超灵敏免疫印迹方法(western blot)用以对复杂蛋白样品进行分析检测,碳纳米管—纸免疫传感器对水中的微囊藻毒素进行检测。一种免疫方法是：基于量子点的超灵敏蛋白检测免疫印迹方法。碲化镉(CdTe)量子点由于它独特的光学及生物特性,被广泛应用于生物化学标记领域。本论文构建了一种基于量子点的超灵敏蛋白检测免疫印迹方法。由于亲和素功能化的聚合量子点的高亲和性及对检测步骤的简化,使得量化免疫印迹方法成为现实。为了制备聚合量子点,首先需要用生物素化的变性牛血清白蛋白(dBSA)包裹量子点,然后通过生物素与亲和素的相互作用,量子点相互偶联生成聚合量子点。经过一系列的修饰,碲化镉量子点的荧光强度显著增强。把聚合量子点作为免疫印迹的标志物,其追踪微量蛋白比传统染料更灵敏。以蛋白A为例,对基于聚合量子点的免疫印迹方法进行验证,检测线性范围为30pg到1.5ng,检测限达0.84pg。最终在聚偏二氟乙烯膜上的荧光信号至少可以保持40min。进行实际样品检测时,回收率达99.9％到103.0％。另一种方法是纸—单壁碳纳米管复合物组成的,简单且高效的传感器。碳纳米管传感器的电阻取决于碳纳米管网络之间的间隙宽度,该方法能很好的满足常规毒素的检测要求。本论文对微囊藻毒素进行了检测并建立了标准曲线,线性范围为1.25nmol／L到10nmol／L。检测限达0.6nmol／L(0.6ng／mL),满足了世界卫生组织关于饮用水中微囊藻毒素含量不超过1ng／mL的检测要求。同时,论文还对该方法与ELISA检测方法进行了比较。更多还原

【Abstract】 Quantum dots and carbon nanotube have been widely used in the field of immunoassay because of their particular speciality and bio-compatibility. In the paper, we researched two detection method based on immunosassy. Ultrasensitive detection of protein by Western blot based on POLY-quantum dot probes were used in micro-protein analyzing, and single walled carbon-nanotubes (SWNTs)-paper sensor were used to detect MC-LR in water.One of the method is Ultrasensitive detection of protein by Western blot based on POLY-quantum dot probes. With the high affinity of avidin-functionalized POLY-QDs and simplification of the detection process, this enabled the quantitative analysis of protein by Western blotting. To prepare the POLY-QDs, CdTe quantum dots were first coated with biotinylated denatured bovine serum albumin (dBSA), and then via the effect of the biotin-avidin system, the biotinylated dBSA-coated QDs which had strong fluorescence were conjugated. With this series of modifications, the fluorescence intensity of CdTe QDs was significantly increased. Using the POLY-QDs as labels, the signal of Western blotting was more sensitive in tracing the protein than traditional dyeing. In the present study, protein A was applied to POLY-QDs-based Western blotting as a model. The linearity of this method was from 30 pg to 1.5 ng, and the sensitivity was up to 0.84 pictogram. The final fluorescence signal on the PVDF membrane was retained for at least 40 min. The recovery of real samples detection is from 99.9% to 103.0%.The other is composites obtained impregnation of papers by SWNTs towards very simple and high-performance biosensors. They utilize the strong dependence of electrical conductivity through nanotubes percolation network on the width of nanotubes-nanotube tunneling gap and can potentially satisfy the requirements for the routine toxin monitoring. In this study, we detected MC-LR in water with the SWNTs-paper biosensor. The method has the linear detection range from 1.25 nmol/L to 10 nmol/L. The detection limit was found to be 0.6 nmol/L (0.6 ng/mL), which satisfies the strected World Health Organization standard for MC-LR content in drinking water (1 ng/mL), and is comparable to the detection limit of traditional ELIS A method of MC-LR detection.更多还原