本文選題：神經(jīng)管缺陷 + Vangl1蛋白��； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：Vangl1蛋白是平面細胞極性信號通路(PCP)的核心蛋白,其表達異常與胚胎發(fā)育不良高度相關(guān),尤其是神經(jīng)管缺陷(NTDs)。由于目前缺乏Vangl1蛋白的痕量檢測方法,因此研究并建立相關(guān)超靈敏檢測分析技術(shù)可能對神經(jīng)管缺陷的精確診斷發(fā)揮潛在的臨床價值。近年來生物傳感器以其具有操作簡單、成本低、檢測限低和靈敏度高等優(yōu)勢,被廣泛應(yīng)用于臨床診斷和生物樣品分析領(lǐng)域,已經(jīng)成為一種生物標志物檢測的理想分析技術(shù)。其工作原理是將生物活性物質(zhì)(核酸、抗體、抗原、酶等)在電極表面發(fā)生分子識別事件轉(zhuǎn)換為聲、光、電物理等可以測定物的信號,再進行分析。本文主要從納米復(fù)合物的制備以及信號放大策略的選擇來構(gòu)建Vangl1蛋白免疫傳感器,并對其分析性能進行研究和檢測,具體研究內(nèi)容和結(jié)果如下:首先采用原位還原方法,在C60表面固載Au Pt NPs形成C60-Au Pt納米復(fù)合材料,作為信標材料,通過催化H2O2產(chǎn)生電信號,實現(xiàn)信號的放大。其次,為進一步提高免疫傳感器的靈敏度,選用PTC-NH2修飾的r GO-TEPA復(fù)合物PTC-NH2-r GO-TEPA作為基底材料,兩者均含有豐富的氨基集團,通過戊二醛(GA)的交聯(lián)作用與抗體結(jié)合,可以固載大量的抗體;同時進行條件優(yōu)化,在最優(yōu)的實驗條件下,所構(gòu)建的免疫傳感器在0.1pg/m L-450ng/m L范圍內(nèi),得到了較好的線性結(jié)果,最低檢測限為0.03pg/m L。最后驗證該免疫傳感器具有良好的穩(wěn)定性與重現(xiàn)性,可用于血液樣品中Vangl1蛋白的檢測。本研究對NTD等出生缺陷的快速臨床分子診斷提供了有意義的實驗依據(jù)。
[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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R714.5;TP212.3

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