基于鲁米诺纳米复合材料的电化学发光免疫传感器的构建及其应用

发布时间：2018-02-09 09:34

本文关键词： 电化学发光免疫传感器鲁米诺纳米复合材料肿瘤标志物　出处：《上海师范大学》2017年硕士论文　论文类型：学位论文

【摘要】：电化学发光技术是发光物质在电极表面通过电子得失产生的化学发光的现象,该技术能够将电能直接转化为光能。近年来,电化学发光技术受到越来越多研究人员的关注,在药物分析、免疫分析、DNA分析、临床分析等科学领域得到广泛运用。新型功能性纳米复合材料由于其特殊的纳米结构、较强的吸附能力、大的比表面积等优势为电化学传感器的研究和应用提供新途径。将其应用到电化学发光免疫传感器的构建中,可大大提高生物分子(如酶、DNA等)的固载量、标记生物分子、催化反应、加快电子传递,增大电流信号等。本论文以将多壁碳纳米管-铂-鲁米诺(MWCNTs-Pt-luminol)作为免疫平台,构建超灵敏的电化学发光免疫传感器检测糖抗原19-9(CA19-9);以MoS2-PEI-Au和Au@BSA两种功能性纳米材料分别作为传感平台和标记鲁米诺(luminol),构建灵敏的电化学发光免疫传感器检测甲胎蛋白(AFP);以luminol标记的Pt@BSA纳米粒子作为包被抗原的固载基质,利用介孔碳球-二茂铁(MCS-Fc)纳米材料并对传感器界面进行修饰,用来检测糖抗原15-3(CA15-3)。本论文主要包括四个方面内容:1.阐述了论文的背景和意义,主要介绍了电化学发光的机理和功能性纳米材料在电化学发光传感器中的应用,电化学免疫传感器在实际生活中的应用等。2.合成MWCNTs-Pt-luminol纳米复合材料,借助紫外可见光谱和透射电镜进行表征。将上述功能性材料进行组装构建了超灵敏的电化学免疫传感器,并用于检测CA19-9。本方法最低检出限为0.000046 U/mL,具有良好的重现性及稳定性,在实际血清样品测定中的结果令人满意。3.胶体金功能化的MoS2纳米片通过静电作用合成MoS2-PEI-Au纳米复合材料固定在电极表面作为传感平台,Au@BSA提供了更大的比表面积用来标记luminol和固定二抗(Ab2),利用抗原与抗体之间的特异性结合构建了灵敏检测甲胎蛋白的夹心型电化学发光免疫传感器。在最佳的实验条件下,该传感器对AFP检测的线性范围从0.0001-200.0ng/m L,检测限为0.00001ng/m L。此外,该传感器对实际样品的检测显示出优良的性能,为其他临床生物标志物的检测提供了巨大的潜力。4.利用具有大的比表面积、良好的导电性以及催化活性的介孔碳球-二茂铁(MCS-Fc)作为基底材料,可以催化H2O2分解,实现电化学发光信号的增强。luminol-Pt@BSA-Ab2作为信号探针,当孵育CA15-3之后,从而成功构建了一个信号放大的夹心型电化学发光免疫传感器,并实现了对CA15-3的超灵敏检测。该免疫传感器具有较宽的线性范围和较低的检测线。在最佳的实验条件下,该传感器对CA15-3检测的线性范围从0.0005-500.0 U/mL,检测限为0.000025 U/m L。此外,该免疫传感器的构建思路为临床上其他生物分子和蛋白质的检测提供了一定的参考价值。
[Abstract]:Electrochemiluminescence (ECL) is a phenomenon of chemiluminescence produced by electroluminescent materials passing through electrons on the electrode surface. This technology can convert electric energy directly into light energy. In recent years, more and more researchers have paid attention to ECL technology. New functional nanocomposites are widely used in the fields of drug analysis, immunoassay, DNA analysis, clinical analysis and so on. The advantages of large specific surface area provide a new way for the research and application of electrochemical sensors. Applying them to the construction of electrochemiluminescence immunosensors can greatly increase the immobilization capacity of biomolecules (such as enzyme DNA) and label biomolecules. In this paper, we take MWCNTs-Pt-luminolol as the immune platform, which is based on multiwalled carbon nanotubes (MWCNTs-Pt-luminolol). A super sensitive electrochemiluminescence immunosensor was constructed to detect sugar antigen 19-9 and CA19-9, and two functional nanomaterials, MoS2-PEI-Au and Au@BSA, were used as sensing platform and labeled luminol, respectively, to construct a sensitive electrochemiluminescence immunosensor for the detection of nail fetal eggs. Luminol labeled Pt@BSA nanoparticles were used as the immobilized matrix for encapsulating the antigen. The mesoporous carbon-ferrocene (MCS-Fc) nanomaterials were used to detect carbohydrate antigens 15-3CCA15-3 by modifying the sensor interface. This paper mainly includes four aspects: 1.The background and significance of the paper are described. The mechanism of electrochemiluminescence and the application of functional nano-materials in electroluminescent sensors, the application of electrochemical immunosensors in practical life, etc. 2. Synthesis of MWCNTs-Pt-luminol nanocomposites are introduced in this paper. Ultraviolet-visible spectroscopy and transmission electron microscope were used to characterize the electrochemical immunosensor. The ultrasensitive electrochemical immunosensor was assembled and used for the detection of CA19-9.The detection limit of the method was 0.000046 UmL, which showed good reproducibility and stability. The results obtained in the actual serum samples are satisfactory .3. Colloidal gold-functionalized MoS2 nanoparticles were synthesized by electrostatic interaction to fix MoS2-PEI-Au nanocomposites on the electrode surface as a sensing platform, providing a larger specific surface area to be used as a sensing platform. A sandwich electrochemiluminescence immunosensor for sensitive detection of alpha-fetoprotein was constructed by labeling luminol and immobilized second antibody Ab2H. Under the optimum experimental conditions, a sandwich electrochemiluminescence immunosensor was constructed for the detection of alpha-fetoprotein by specific binding between antigens and antibodies. The linear range of the sensor for AFP detection is 0.0001-200.0ngr / mL, and the detection limit is 0.00001ngr / mL. in addition, the sensor shows excellent performance for the detection of real samples. It provides a great potential for the detection of other clinical biomarkers. Using MCS-Fc with large specific surface area, good electrical conductivity and catalytic activity as substrate material, it can catalyze the decomposition of H _ 2O _ 2. The enhancement of electrochemiluminescence signal. Luminol-Ptr BSA-Ab2 was used as a signal probe. After incubating CA15-3, a sandwich electroluminescent immunosensor with amplified signal was successfully constructed. The sensitivity detection of CA15-3 is realized. The immunosensor has a wide linear range and a lower detection line. Under the optimum experimental conditions, the linear range of detection for CA15-3 is 0.0005-500.0 UmLand the detection limit is 0.000025 U / mL. The construction of the immunosensor provides a certain reference value for the detection of other biomolecules and proteins in clinic.
【学位授予单位】：上海师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.1

【参考文献】