SERS活性磁性纳米复合材料制备及其在肿瘤蛋白标志物检测中的应用研究

发布时间：2018-04-10 14:49

本文选题：表面增强拉曼散射 + 磁性纳米颗粒　；参考：《南京邮电大学》2015年硕士论文

【摘要】：随着现代医学的发展,大量研究证明肿瘤的产生和发展过程中都伴随着一些特定蛋白质的出现或者含量变化,检测肿瘤标志蛋白及其含量有望对肿瘤实现早期诊断。表面增强拉曼散射(SERS)用于蛋白质检测是蛋白质检测学与SERS技术相结合的新型检测技术,在生物医学检测领域表现出巨大的应用潜力。基于三明治结构的SERS免疫检测是常用的一种基于SERS技术的蛋白质检测方法,基本组成部分有免疫检测基底、待检测蛋白、SERS免疫检测探针。近年来,磁性纳米材料在生物样品的分离提纯方面表现出了突出的优势,是理想的免疫基底材料。本论文中我们可控制备了Fe3O4@SiO2@Au核壳结构这一兼具磁性和SERS活性的纳米材料,并进一步构建得到免疫检测基底,结合花状金纳米颗粒构建的SERS免疫探针,实现了对肿瘤标志蛋白CEA、NSE的高灵敏联合免疫检测。主要研究内容如下:(1)可控制备复合SERS增强性能的磁性纳米材料首先采用共沉积法制备了Fe3O4磁性纳米颗粒,随后对纳米颗粒进行二氧化硅包壳,制备Fe3O4@SiO2核壳型纳米结构,通过控制正硅酸乙酯的加入剂量调控了Fe3O4@SiO2的壳厚和包覆磁性颗粒粒子数,并进一步吸附金种子后生长了一层SERS活性金纳米壳,制备得到复合SERS增强性能的磁性纳米材料Fe3O4@SiO2@Au。(2)基于SERS活性磁性纳米颗粒开展肿瘤标志蛋白检测选择磁性和结构俱佳、SERS性能优异的Fe3O4@SiO2@Au,经表面修饰特定抗体构建得到用于捕获待检测蛋白的免疫检测基底,同时制备了具有优异SERS增强性能的花状金纳米颗粒并构建了SERS免疫检测探针。免疫识别后形成三明治结构复合物,利用外加磁场,实施检测样品的分离与富集,分别对human-IgG和肿瘤标志蛋白CEA实现了高灵敏、特异性检测。(3)肿瘤标志蛋白联合检测为实现双元肿瘤蛋白标志物的联合检测,我们基于Fe3O4@SiO2@Au材料,设计并构建了能够同时特异性捕获肿瘤标志物CEA和NSE的磁性免疫基底,并分别构建了用于检测CEA和NSE的高灵敏SERS探针,开展并实现了对两种肿瘤标志蛋白的低浓度、特异性联合检测。
[Abstract]:With the development of modern medicine, a large number of studies have proved that the emergence and development of tumors are accompanied by the emergence or content of some specific proteins, the detection of tumor markers and their content is expected to achieve early diagnosis of tumors.Surface enhanced Raman scattering (SERS) for protein detection is a new detection technique combining protein detection with SERS technology. It has great potential in biomedical detection.Sandwich structure based SERS immunoassay is a commonly used protein detection method based on SERS technology.In recent years, magnetic nanomaterials have shown outstanding advantages in the separation and purification of biological samples and are ideal immune substrate materials.In this thesis, we can control the Fe3O4@SiO2@Au core-shell structure, which is a magnetic and SERS active nano-material, and further construct the immunosensory substrate and construct the SERS immune probe combined with the flower-like gold nanoparticles.A highly sensitive combined immunoassay for tumor marker CEANSE was achieved.The main contents of the study are as follows: (1) the magnetic nanocrystalline materials which can control the properties of composite SERS were prepared by co-deposition method, and then Fe3O4@SiO2 core-shell nanostructures were prepared by encapsulating the nanoparticles with silica.By controlling the dosage of ethyl orthosilicate, the shell thickness and the number of particles coated with magnetic particles of Fe3O4@SiO2 were regulated, and a layer of SERS active gold nanoparticles was grown after further adsorption of gold seeds.To capture the immunoassay substrate of the protein to be tested,At the same time, flower-like gold nanoparticles with excellent SERS enhancement properties were prepared and SERS immunoassay probes were constructed.The sandwich structure complex was formed after immunological recognition. The sample was separated and enriched by external magnetic field. The sandwich structure complex was highly sensitive to human-IgG and tumor marker protein CEA, respectively.A highly sensitive SERS probe was constructed for the detection of CEA and NSE, and the low concentration and specificity detection of the two tumor marker proteins was carried out and realized.
【学位授予单位】：南京邮电大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;O657.37

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