一种基于磁性石墨烯的纸基电化学发光夹心免疫分析方法

发布时间：2018-10-31 11:45

【摘要】：基于电化学发光及磁悬浮免疫分析策略,结合磁性石墨烯独特的物理化学特性以及纸基电极价格低廉、样品用量少的优势,建立了一种新型免疫分析方法。以人免疫球蛋白G(IgG)为分析物,采用1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺/N-羟基硫代琥珀酰亚胺(EDC/NHS)法将一抗(Ab_1,捕获抗体)固定在磁性石墨烯上,通过直接标记法进行二抗(Ab_2,信号抗体)的电化学发光试剂标记,采用磁悬浮夹心免疫技术最大程度减少非特异性吸附,通过纸基电化学发光检测技术测定目标物的浓度。考察了捕获抗体及信号抗体的固定(标记)效果,发现采用的磁性石墨烯不仅提高了免疫物质的负载量,还可以促进电子传递,构建的磁悬浮纸基电化学发光夹心免疫分析法的电化学发光响应峰面积在0.32~1000 ng/mL浓度范围内,与IgG浓度对数值呈良好的线性关系,检出限为6.4 pg/mL。本方法可实现IgG的定量检测,在低成本、快速免疫检测领域有一定的应用前景。
[Abstract]:Based on the strategies of electrochemiluminescence and maglev immunoassay, a novel immunoassay method was established based on the unique physical and chemical properties of magnetic graphene and the advantages of low price of paper based electrode and less sample consumption. Using human immunoglobulin G (IgG) as the analytical material, the first antibody (Ab_1,) was determined by 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide / N-hydroxythiosuccinimide (EDC/NHS) method. The captured antibody was immobilized on the magnetic graphene, and the electrochemiluminescence (ECL) labeling of the second antibody (Ab_2, signal antibody) was carried out by direct labeling method. The magnetic levitation sandwich immunoassay was used to minimize the nonspecific adsorption. The concentration of the target was determined by paper-based electrochemiluminescence (ECL). The immobilization (labeling) effect of capture antibody and signal antibody was investigated. It was found that magnetic graphene not only increased the load of immune material, but also promoted electron transfer. The electrochemical luminescence response peak area of magnetic levitation paper based electrochemiluminescence sandwich immunoassay was within the concentration range of 0.32 ~ 1000 ng/mL, which showed a good linear relationship with the logarithmic value of IgG concentration, and the detection limit was 6.4 pg/mL.. This method can be used for quantitative detection of IgG, and has a certain application prospect in the field of low cost and fast immune detection.
【作者单位】：广州军区广州总医院医学实验科;
【基金】：国家自然科学基金(No.81401752) 广州市科技计划(No.201607010121) 军队后勤保障项目(No.CGZ16J002)资助~~
【分类号】：O657.1

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