基于纳米复合材料和蛋白质的生物传感新方法及其应用—葡萄糖及呕吐毒素快速检测

发布时间：2018-01-03 12:13

本文关键词：基于纳米复合材料和蛋白质的生物传感新方法及其应用—葡萄糖及呕吐毒素快速检测　出处：《合肥工业大学》2016年硕士论文　论文类型：学位论文

【摘要】：电化学分析是基于物质的电化学性质及其变化规律进行分析的仪器分析方法,仪器易于微小型化、方法简单,灵敏度高,选择性强以及灵活便利等优点而被广泛应用于食品分析,医疗及环境检测等研究。纳米材料具有丰富的表面活性位点,良好的生物相容性,较大的比表面积,较高的活性与催化效率,以及较强吸附能力等特点。在电化学生物传感器领域,纳米材料是修饰电极以及固定生物分子的理想材料,其自身优异性能可以显著地提高生物传感器的检测性能。本文基于这一研究背景,以葡萄糖和呕吐毒素(DON)为检测对象,利用石墨烯和金属纳米粒子等材料修饰玻碳电极(GCE),构建了性能优良的蛋白质电化学生物传感器,如葡萄糖氧化酶(GOx)生物传感器和呕吐毒素(DON)抗原-抗体免疫传感器,建立了对葡萄糖和呕吐毒素的快速传感新方法。本文研究内容主要包含以下三个部分：1.纳米复合材料的制备及表征利用壳聚糖(CS)作为还原剂和稳定剂一步法合成了纳米金-还原氧化石墨烯(rGO-AuNPs)纳米复合材料。通过红外光谱,紫外-可见光光谱和场发射透射电子显微镜表征其结构及表面形态。可以证明AuNPs成功负载在还原氧化石墨烯片层上。相比于裸电极以及仅由rGO或AuNPs单独修饰玻碳电极(GCE),rGO-AuNPs修饰GCE具有更好的导电性能。利用十二烷基硫酸钠作为结构导向剂,制备了具有优异离子交换性能的磷酸钛纳米球(TiP纳米球)。然后将Ag+负载到磷酸钛纳米球上,再将Ag+还原为银纳米粒子形成AgNPs@TiP纳米复合材料。通过紫外-可见光光谱、X射线光电子能谱、场发射扫描电子显微镜和X射线能谱仪表征了AgNPs@TiP的结构及表面形态。可以证明AgNPs成功负载在TiP纳米球上,AgNPs@TiP的平均粒径约55nm。因其具有优异的离子交换能力和丰富的活性位点,AgNPs@TiP对蛋白质传感器研究具有潜在的应用价值。2.葡萄糖氧化酶-纳米复合材料修饰电极安培法检测葡萄糖使用rGO-AuNPs修饰GCE后并固定GOx,构建GOx/rGO-AuNPs/GCE学生物传感器,实现了GOx在GCE的直接电化学。通过循环伏安法和计时电流法系统地研究了酶的用量、磷酸盐缓冲溶液(PBS)的pH值等制备工艺对葡萄糖氧化酶生物传感器的影响。GOx在传感器上的表观米氏常数KmaPP为0.497,对0.010～0.88 mM范围内的葡萄糖具有优异的检测效果,检测限低至0.43μM(信噪比S/N=3),灵敏度高达22.54 μAmM-1 cm-2。该方法与葡萄糖试剂盒测定实际饮料样品的测定结果相一致。3.纳米银修饰磷酸钛纳米球-呕吐毒素免疫传感器伏安法快速检测呕吐毒素采用AgNPs@TiP作为信号放大标签,在玻碳电极上修饰电还原氧化石墨烯和壳聚糖用于固定呕吐毒素抗体。依靠游离的呕吐毒素与结合AgNPs@TiP的呕吐毒素竞争免疫实现对呕吐毒素的超灵敏检测。信号标签中的纳米银与底液里的KC1通过差分脉冲伏安法产生一个尖锐的信号峰,峰大小与抗原浓度相对应。该免疫传感器对呕吐毒素的检测范围宽至0.10～1000ng mL-1,检测限为0.059 ng mL-1(信噪比S/N=3)。此外,还原氧化石墨烯优异的导电性和壳聚糖良好的生物相容性保证了免疫传感器具有优秀的稳定性和再生性。
[Abstract]:Electrochemical analysis instrument of electrochemical properties and change of material based on the analysis method, the instrument is easy to be miniaturized, the method is simple, high sensitivity, strong selectivity and flexible and convenient advantages and is widely used in food analysis, medical research and environmental testing. Nano materials with surface active sites rich, good compatibility the biological, large surface area, high catalytic activity and efficiency, and strong adsorption ability. In the field of electrochemical biosensor, nanomaterials are ideal materials modified electrodes and fixed biological molecules, its excellent performance can significantly improve the detection performance of the biosensor. In this paper, based on this research background. With glucose and deoxynivalenol (DON) for the detection of objects, using materials such as graphene and metal nanoparticles modified glassy carbon electrode (GCE), construct a Protein electrochemical biosensor is excellent, such as glucose oxidase (GOx) biosensor and deoxynivalenol (DON) antigen antibody immune sensor, a new method for rapid sensing of glucose and vomitoxin. The contents of this paper mainly includes the following three parts: 1. nano composites were prepared and characterized by chitosan (CS) as a reducing agent and stabilizer was synthesized by the reaction of gold nanoparticles reduced graphene oxide (rGO-AuNPs) nano composite material. By infrared spectroscopy, UV Vis spectroscopy and field emission transmission electron microscopy to characterize the structure and surface morphology. It can be proved that the AuNPs successfully loaded on the reduced graphene oxide layers on. On the bare electrode and only by rGO or AuNPs alone modified glassy carbon electrode (GCE), conductive properties of rGO-AuNPs modified GCE has better. By using twelve sodium dodecyl sulfate as structure directing agent system Titanium phosphate nanoparticles have excellent ion exchange properties were synthesized (TiP spheres). Then Ag+ was loaded into the nano titanium phosphate on the ball, then Ag+ reduced to silver nanoparticles formation of AgNPs@TiP nanocomposites. By UV visible spectroscopy, X ray photoelectron spectroscopy, field emission scanning electron microscope and X ray energy dispersive X-ray spectrometer structure and surface morphology of AgNPs@TiP. It can be proved that the AgNPs successfully loaded on TiP nanoparticles, AgNPs@TiP average particle size of about 55nm. because of its excellent ion exchange capacity and abundant active sites, AgNPs@TiP has the potential application value of detection of glucose glucose oxidase.2. nanocomposites modified electrode using amperometric method rGO-AuNPs GCE and GOx on protein fixed sensor research, construction of biological sensor GOx/rGO-AuNPs/GCE, implemented in GOx GCE by direct electrochemical cyclic voltammetry. Chronoamperometry and systematic study of the enzyme dosage, phosphate buffer solution (PBS) of the pH value and the influence of the preparation process of glucose oxidase biosensor in the.GOx sensor on the apparent Michaelis constant KmaPP was 0.497, 0.010 to 0.88 mM within the scope of the glucose detection results is excellent, low detection limit to 0.43 M (signal-to-noise ratio S/N=3), determination of the results of the sensitivity up to 22.54 AmM-1 cm-2. glucose assay kit and the method of actual beverage samples consistent with.3. nano silver modified titanium phosphate nanoparticles - vomitoxin immunosensor for rapid detection of deoxynivalenol by voltammetry using AgNPs@TiP as signal amplification label, modified electrochemical reduction oxidation graphene and chitosan for fixing the emetic toxin antibody on glassy carbon electrode. Rely on mycotoxin free and bound voitoxin AgNPs@TiP for the competitive immune deoxvnivalenol ultra sensitive Detection. Silver nanoparticles and base solution in the KC1 tag signal by differential pulse voltammetry to produce a sharp peak, peak size and corresponding antigen concentration. The immune sensor detection range of the deoxvnivalenol width to 0.10 ~ 1000ng mL-1, the detection limit is 0.059 ng mL-1 (SNR S/N=3). In addition, the reduction of conductivity of graphene oxide and chitosan with excellent biocompatibility to ensure the immune sensor has excellent stability and reproducibility.

【学位授予单位】：合肥工业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：O657.1;TP212

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