基于Poly（T）DNA和BSA模板铜纳米簇的荧光生传感新方法研究

发布时间：2018-05-31 16:12

本文选题：荧光铜纳米簇 + 二氧化锰纳米片　；参考：《信阳师范学院》2017年硕士论文

【摘要】：荧光金属纳米团簇具有尺寸小、易于合成、较强的抗光漂白性、良好的生物相容性等优点,是一种新型的纳米材料。近年来,由于荧光铜纳米簇具有成本低、水溶性好、光学性能稳定、可调的荧光发射等特点,在生物分析、生物成像、医疗应用、环境监测等领域引起了广泛的研究兴趣。本文利用Poly(T)DNA和牛血清白蛋白作为合成模板,合成了不同荧光发射波长的荧光铜纳米簇,并以此作为荧光信号探针,检测生物小分子和巯基化合物。此外,蛋白质为模板的荧光铜纳米簇可以与二氧化锰纳米片形成复合物,开发了一种新型、灵敏、无标记的传感新方法,用于蛋白质的检测。主要研究内容如下:(1)利用曲酸阻止Poly(T)DNA模板的荧光铜纳米簇的形成,构建了一种简单,快速,非标记型的荧光传感新方法并用于曲酸的灵敏检测。荧光铜纳米簇以Poly(T)DNA作为模板,以抗坏血酸为还原剂,在室温下5分钟内合成。当目标曲酸存在时,由于Cu~(2+)和曲酸形成稳定的络合物,有效地阻止了Poly(T)DNA模板荧光铜纳米簇形成,从而导致体系荧光信号减弱。在优化的条件下,该方法对曲酸检测的线性范围为0.1~300μmol/L,检出限为10 nmol/L。同时,该方法用于食品样品中曲酸含量的测定,RSD范围在2.7%~4.2%,表明该方法是可行的。(2)基于阻碍半胱氨酸(Cys)猝灭Poly(T)DNA模板的荧光铜纳米簇的荧光,发展了一种快速、灵敏、无标记的荧光生物传感策略用于Hg~(2+)的检测。在Cys存在的情况下,Cys与铜纳米簇之间形成稳定的Cu-S键,从而有效地猝灭铜纳米簇的荧光信号。当体系加入Hg~(2+)后,Cys与Hg~(2+)形成了更稳定的Hg-S键,导致Cys远离了荧光铜纳米簇的表面,体系的荧光强度得到恢复。该方法实现了对目标Hg~(2+)的灵敏检测,线性范围为0.5~100 nmol/L,检出限为0.1 nmol/L,用于实际样品检测,RSD在2.36%~3.12%,表明该方法可用于实际样品的检测。(3)基于MnO_2纳米片-荧光铜纳米簇复合物,建立了一种新型的荧光传感策略用于谷胱甘肽的高灵敏、高选择性检测。荧光铜纳米簇以牛血清白蛋白为模板,通过简便的一步化学还原法制得。当MnO_2纳米片-荧光铜纳米簇复合物形成时,铜纳米簇的荧光信号能够通过荧光能量共振转移被MnO_2纳米片有效地猝灭。当加入目标谷胱甘肽时,MnO_2纳米片能够被还原生成Mn2+,从而破坏了MnO_2纳米片,使MnO_2纳米片引起的荧光猝灭效应得到抑制,最终体系的荧光信号得到恢复。该方法的线性检测范围为0~300μmol/L,检出限为100 nmol/L,用于实际样品检测,RSD范围3.3%~4.3%,同时具有较好的选择性。结果表明,该方法具有简单,快速,成本低,易于制备等优点。(4)利用MnO_2纳米片-荧光铜纳米簇复合物作为荧光探针,基于H_2O_2诱导破坏猝灭剂MnO_2纳米片,开发了一种灵敏的荧光生物传感新方法用于葡萄糖的测定。当H_2O_2存在时,MnO_2被还原生成Mn2+,铜纳米簇从MnO_2纳米片表面释放出来,体系的荧光信号得到恢复。另一方面,葡萄糖在葡萄糖氧化酶存在条件下,被催化氧化生成葡萄糖酸和H_2O_2,因此,通过酶催化产生的H_2O_2导致的荧光信号,可以间接地检测葡萄糖的浓度。该方法的线性检测范围为1~200μmol/L,检出限为100 nmol/L。同时,RSD在2.9%~4.2%,表明该方法可用于人血清样品中葡萄糖含量的测定。
[Abstract]:The fluorescent metal nanoclusters have the advantages of small size, easy synthesis, strong anti photobleaching, good biocompatibility and so on. It is a new kind of nano material. In recent years, the fluorescent copper nanoclusters have the characteristics of low cost, good water solubility, stable optical properties, adjustable fluorescein and so on, in biological analysis, bioimaging, medical applications, and so on. In this paper, Poly (T) DNA and bovine serum albumin (BSA) are used as a synthetic template to synthesize fluorescent copper nanoclusters with different fluorescence emission wavelengths, which are used as a fluorescent probe to detect small biological molecules and sulfhydryl compounds. In addition, the fluorescent copper nanoclusters of protein as a template can be used as a template. A new, sensitive and unmarked new sensing method was developed for the detection of protein. The main contents of this study are as follows: (1) a simple, fast, unlabeled new fluorescence sensing method was constructed and used in the spirit of kojic acid by using kojic acid to prevent the formation of Poly (T) DNA template. Poly (T) DNA as a template and ascorbic acid as a reductant and synthesized in 5 minutes at room temperature. When target ascorbic acid exists, Cu~ (2+) and kojic acid form a stable complex, which effectively prevents the formation of Poly (T) DNA template fluorescent copper nanoclusters, resulting in a weakening of the fluorescence signal of the system. At the same time, the linear range of the method is 0.1~300 mu mol/L and the detection limit is 10 nmol/L.. The method is used to determine the content of kojic acid in food samples. The RSD range is 2.7%~4.2%, indicating that the method is feasible. (2) a rapid development is developed based on the fluorescence of the fluorescent copper nanoclusters that hinder the quenching of Poly (T) DNA template with cysteine (Cys). A sensitive, unmarked fluorescence biosensor strategy is used for the detection of Hg~ (2+). In the presence of Cys, a stable Cu-S bond between Cys and copper nanoclusters is formed, which effectively quenches the fluorescence signals of the copper nanoclusters. When Hg~ (2+) is added to the system, Cys and Hg~ (2+) form a more stable Hg-S bond, which leads to Cys away from the table of the fluorescent copper nanoclusters. The fluorescence intensity of the system is restored. The method realizes the sensitive detection of the target Hg~ (2+). The linear range is 0.5~100 nmol/L, the detection limit is 0.1 nmol/L. The method is used for the actual sample detection and RSD in 2.36%~3.12%. It shows that the method can be used for the detection of actual samples. (3) a kind of MnO_2 nanoscale copper nanocluster complex is established. The new fluorescence sensing strategy is used for high sensitivity and high selectivity of glutathione. The fluorescent copper nanocluster is prepared by a simple one-step chemical reduction method using bovine serum albumin as a template. When the MnO_2 nanoscale copper nanocluster complex is formed, the fluorescence signal of the copper nanocluster can be transferred by MnO_2 to the fluorescence energy resonance. MnO_2 nanoscale can be reduced to Mn2+ when the target glutathione is added. The MnO_2 nanoscale is destroyed and the fluorescence quenching effect caused by the MnO_2 nanoscale is suppressed. The fluorescence signal of the final system is restored. The linear detection of this method is 0~300 micron mol/L, the detection limit is 100 nmol/L, used for real time. The results show that the method has the advantages of simple, rapid, low cost, easy to prepare and so on. (4) a sensitive fluorescent biological transmission is developed by using MnO_2 nanoscale fluorescent copper nanocluster complex as a fluorescent probe and H_2O_2 induced sudden destruction agent MnO_2 nanoscale based on H_2O_2. A new sense method is used for the determination of glucose. When H_2O_2 exists, MnO_2 is reduced to Mn2+, and the copper nanocluster is released from the surface of MnO_2 nanoscale. The fluorescence signal of the system is restored. On the other hand, glucose is catalyzed to produce gluconic acid and H_2O_2 under the condition of glucose oxidase. Therefore, the H_2 produced by enzyme catalyzed. The fluorescence signal caused by O_2 can be used to detect the concentration of glucose indirectly. The linear detection range of this method is 1~200 mol/L, the detection limit is 100 nmol/L., and RSD is 2.9%~4.2%. It shows that the method can be used for the determination of glucose content in human serum samples.
【学位授予单位】：信阳师范学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

【参考文献】