血蓝蛋白及半乳糖氧化酶复合膜电化学传感

发布时间：2018-06-04 18:40

本文选题：半乳糖氧化酶 + 石墨烯　；参考：《湘潭大学》2017年硕士论文

【摘要】：由于第三代酶生物传感器在医疗和环境方面的良好的应用前景,与第三代生物传感器的相关课题引起了科研人员的巨大兴趣。21世纪开始的阶段,材料科学方面取得的进步,给第三代生物传感器带来了新的机遇。纳米材料以及导电聚合物,改进了第三代生物传感器的一些相关性质。其中,石墨烯的导电性比以往的纳米材料有所提高,比表面积较大,适合生物分子的固定。有助于生物分子活性的保留。本文制作了三种性能良好的的第三代生物传感器,利用功能化石墨烯等材料固定血蓝蛋白,半乳糖氧化酶,对所制备的生物传感器进行了研究。主要内容为:使聚乙二醇乙二酸脂(Poly(ethylene succinate),PES)与氧化石墨烯(GO)结合。对氧化石墨烯(GO)在-0.60 V的电压下进行电还原处理,获得还原氧化石墨烯与PES复合材料,首次利用这种RGO/PES复合膜固载了血蓝蛋白(hemocyanin,HC),形成了生物复合膜RGO/PES/HC,我们用扫描电子显微镜(SEM)表征这种生物复合膜的表面形貌,电化学交流阻抗(EIS)以及循环伏安法(CV)等电化学测试方法也被用于测试本工作制备的RGO/PES/HC修饰电极的电化学相关性质。获得了在不同pH条件和扫速的情况下的循环伏安图,通过分析循环伏安图来研究纳米膜复合膜的电子传递过程。同时,用CV和微分脉冲伏安法(DPV)来研究本工作中制备的RGO/PES/HC修饰电极对双酚A(Bisphenol A,BPA)的催化情况。实验结果表明,制作的生物传感器对BPA的检测的线性范围为8.0×10-8M到5.5×10-5M,检出限为3.4×10-8 M,并利用本工作制备的生物传感器对塑料材料产品中的双酚A进行了实样检测,回收率在可接受范围之内,结果仍然是让人满意的。利用原位电化学聚合的方法,合成了导电聚合物聚苯胺(polyaniline,PANI)/RGO复合膜,同时在原位生成聚苯胺的过程中,利用静电作用的原理,将半乳糖氧化酶(GAO)固定在RGO/PANI纳米复合材料内,利用交流阻抗(EIS)和循环伏安法(CV)研究了不同修饰电极的电化学性质。并且利用安培法,研究了复合膜对于半乳糖的催化行为,通过计算得到的检测限为9.0×10-6 M。同时对实验条件进行了优化,我们发现在PH=7.0,电位为-0.50 V时,获得最好的检测效果。在测试所制备的生物传感器的抗干扰能力方面,检测了常见干扰物的干扰情况,情况令人满意,将制备好的修饰电极在冰箱中保存15天后,仍然有92%的响应电流。对稳定性和重现性进行测试,发现本工作构建的酶电化学生物传感器的稳定性和重现性良好。利用聚甲基丙烯酸甲酯(polymethyl methacrylate,PMMA)功能化氧化石墨烯(GO),将半乳糖氧化酶(Galactose oxidase,GAO)固定在GO/PMMA纳米复合材料之内,用SEM研究了GO/PMMA/GAO生物复合膜的表面形貌,进一步利用CV研究了生物复合膜的电化学行为,并在不同的pH和扫速下进行了循环伏安测试,研究了电极表面的电化学行为,同时用时间电流法(i-t)研究了传感器对半乳糖和二羟基丙酮(1,3-Dihydroxyacetone,DHA)的催化行为,其中对半乳糖催化的线性范围为7.0×10-6 M到2.4×10-4 M,对二羟基丙酮的线性检测范围为8.0×10-6 M到4.5×10-4 M检出限7.0×10-6 M,并且,通过重现性实验说明GO/PMMA/GAO生物传感器具有良好的重现性。
[Abstract]:Due to the good application prospect of the third generation enzyme biosensor in the medical and environmental aspects, the related topics of the third generation biosensors have aroused the great interest of the researchers in the.21 century. The progress in material science has brought new opportunities to the third generation biosensors. Some of the related properties of the third generation biosensors are improved. Among them, the conductivity of graphene is higher than that of the previous nanomaterials, and the specific surface area is larger. It is suitable for the immobilization of biomolecules. It is helpful for the retention of biological molecules. In this paper, three kinds of third generation biosensors with good performance have been made, using functional fossils and so on. The materials are immobilized on hemocyanin and galactose oxidase, and the biosensors are studied. The main contents are: combining Poly (ethylene succinate) and PES with graphene oxide (GO). The electrochemical reduction of graphene oxide (GO) under the electric pressure of -0.60 V is carried out, and the reduction of graphene oxide and PES is obtained. Materials, the hemocyanin (HC) was immobilized by the RGO/PES composite membrane for the first time, and the biological composite membrane RGO/PES/HC was formed. We used scanning electron microscopy (SEM) to characterize the surface morphology of the composite membrane. Electrochemical impedance (EIS) and cyclic voltammetry (CV) methods were also used to test the preparation of this work. The electrochemical properties of the RGO/PES/HC modified electrode have been obtained. The cyclic voltammograms are obtained at different pH conditions and scavenging conditions. By analyzing the cyclic voltammograms, the electron transfer process of the nanocomposite membranes is studied. At the same time, the RGO/PES/HC modified electrodes prepared in this work are studied by CV and differential pulse voltammetry (DPV) to the bisphenol A (Bis). The experimental results show that the linear range of the detection of BPA by biosensor is 8 * 10-8M to 5.5 x 10-5M, the detection limit is 3.4 * 10-8 M, and the biological sensor prepared by this work is used to detect the bisphenol A in the plastic material, and the recovery rate is within acceptable range, and the result is still in the phenol. It is satisfactory. The polyaniline (PANI) /RGO composite membrane was synthesized by in situ electrochemical polymerization. In the process of in situ formation of polyaniline, the galactose oxidase (GAO) was immobilized in RGO/PANI nanocomposites by electrostatic action, and the AC impedance (EIS) and cyclic voltammetry (CV) method were used. CV) the electrochemical properties of different modified electrodes were studied. The catalytic behavior of the composite membrane for galactose was studied by amperometric method. The detection limit of 9 x 10-6 M. was calculated and the experimental conditions were optimized. We found that the best detection effect was obtained at PH=7.0, when the potential was -0.50 V. With respect to the anti-interference ability of the sensor, the interference of common interferers is detected. The situation is satisfactory. The prepared modified electrode still has 92% response current for 15 days in the refrigerator. The stability and reproducibility are tested. It is found that the stability and reproducibility of the enzyme electrochemistry biosensor constructed in this work are good. With polymethyl methacrylate (PMMA) functionalized graphene oxide (GO), galactose oxidase (Galactose oxidase, GAO) was immobilized in GO/PMMA nanocomposite. The surface morphology of GO/PMMA/GAO biological composite membrane was studied by SEM, and the electrochemical behavior of the biological composite membrane was studied by CV. At the same pH and sweep speed, cyclic voltammetry was carried out to study the electrochemical behavior of the electrode surface. At the same time, the catalytic behavior of the sensor to galactose and two hydroxyacetone (1,3-Dihydroxyacetone, DHA) was studied by time current method (I-T). The linear range of the catalysis of galactose was 7 x 10-6 M to 2.4 x 10-4 M, to the line of two hydroxy acetone. The detection limit is 8 * 10-6 M to 4.5 * 10-4 M detection limit 7 * 10-6 M, and reproducibility test shows that GO/PMMA/GAO biosensor has good reproducibility.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.1;TB33

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