神经管缺陷相关蛋白Vangl1的新型免疫传感器构建

发布时间：2018-05-29 05:53

本文选题：神经管缺陷 + Vangl1蛋白　；参考：《重庆医科大学》2017年硕士论文

【摘要】：Vangl1蛋白是平面细胞极性信号通路(PCP)的核心蛋白,其表达异常与胚胎发育不良高度相关,尤其是神经管缺陷(NTDs)。由于目前缺乏Vangl1蛋白的痕量检测方法,因此研究并建立相关超灵敏检测分析技术可能对神经管缺陷的精确诊断发挥潜在的临床价值。近年来生物传感器以其具有操作简单、成本低、检测限低和灵敏度高等优势,被广泛应用于临床诊断和生物样品分析领域,已经成为一种生物标志物检测的理想分析技术。其工作原理是将生物活性物质(核酸、抗体、抗原、酶等)在电极表面发生分子识别事件转换为声、光、电物理等可以测定物的信号,再进行分析。本文主要从纳米复合物的制备以及信号放大策略的选择来构建Vangl1蛋白免疫传感器,并对其分析性能进行研究和检测,具体研究内容和结果如下:首先采用原位还原方法,在C60表面固载Au Pt NPs形成C60-Au Pt纳米复合材料,作为信标材料,通过催化H2O2产生电信号,实现信号的放大。其次,为进一步提高免疫传感器的灵敏度,选用PTC-NH2修饰的r GO-TEPA复合物PTC-NH2-r GO-TEPA作为基底材料,两者均含有丰富的氨基集团,通过戊二醛(GA)的交联作用与抗体结合,可以固载大量的抗体;同时进行条件优化,在最优的实验条件下,所构建的免疫传感器在0.1pg/m L-450ng/m L范围内,得到了较好的线性结果,最低检测限为0.03pg/m L。最后验证该免疫传感器具有良好的稳定性与重现性,可用于血液样品中Vangl1蛋白的检测。本研究对NTD等出生缺陷的快速临床分子诊断提供了有意义的实验依据。
[Abstract]:Vangl1 protein is the core protein of planar cell polarity signal pathway. The abnormal expression of Vangl1 protein is highly correlated with embryonic dysplasia, especially the neural tube defect. Due to the lack of trace detection methods for Vangl1 protein, the study and establishment of the relevant hypersensitive detection and analysis technology may play a potential clinical value in the accurate diagnosis of neural tube defects. In recent years, biosensor has been widely used in the field of clinical diagnosis and biological sample analysis because of its advantages of simple operation, low cost, low detection limit and high sensitivity. It has become an ideal analysis technology for biomarker detection. Its working principle is to convert the molecular recognition events of bioactive substances (nucleic acid, antibody, antigen, enzyme, etc.) on the electrode surface into the signals of acoustics, light, electrophysics and so on, and then analyze the signals of bioactive substances (such as nucleic acid, antibodies, antigens, enzymes, etc.). In this paper, Vangl1 protein immunosensor was constructed from the preparation of nanocomposites and the selection of signal amplification strategy, and its analytical properties were studied and tested. The specific research contents and results are as follows: firstly, in situ reduction method was used. Au Pt NPs was immobilized on the surface of C60 to form C60-Au Pt nanocomposites, which were used as beacon materials to produce electrical signals by catalyzing H2O2 to amplify the signals. Secondly, in order to improve the sensitivity of the immunosensor, the r GO-TEPA complex PTC-NH2-r GO-TEPA modified by PTC-NH2 was selected as the substrate material. Both of them contain abundant amino groups and bind to antibodies through the crosslinking of glutaraldehyde (Glutaraldehyde). At the same time, under the optimal experimental conditions, the constructed immunosensor obtained good linear results in the range of 0.1pg/m L-450ng/m L, and the lowest detection limit was 0.03pg/m L. Finally, it was proved that the immunosensor had good stability and reproducibility, and could be used for the detection of Vangl1 protein in blood samples. This study provides a significant experimental basis for rapid clinical molecular diagnosis of birth defects such as NTD.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.5;TP212.3

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