基于DNA适配体的C反应蛋白SERS磁性生物传感器的研究  

DNA aptamer-based SERS magnetic biosensor for C-reactive protein

作　　者：张炎 李睿 李利军[1,2] ZHANG Yan;LI Rui;LI Lijun(Guangxi Key Laboratory of Green Processing of Sugar Resources(Guangxi University of Science and Technology),Liuzhou 545006,China;School of Biological and Chemical Engineering,Guangxi University of Science and Technology,Liuzhou 545006,China)

机构地区：[1]广西糖资源绿色加工重点实验室(广西科技大学),广西柳州545006 [2]广西科技大学生物与化学工程学院,广西柳州545006

出　　处：《广西科技大学学报》2022年第3期81-90,106,共11页Journal of Guangxi University of Science and Technology

基　　金：广西自然科学基金项目(2019GXNSFDA245025);国家自然科学基金项目(81860633)资助。

摘　　要：利用C反应蛋白(C-reactive protein,CRP)的巯基化适配体修饰金壳磁性颗粒制备了磁性捕获基底Fe3O4@PEI@Au@Aptamer-SH;以拉曼报告分子对巯基苯甲酸(4-MBA)修饰银纳米粒子,并通过酰胺键偶联CRP的氨基化适配体,制备了信号探针Ag@4-MBA@Aptamer-NH_(2)。磁性捕获基底、抗原CRP以及SERS探针形成三明治夹心复合物,磁分离富集后,实现对CRP的SERS检测。在0.1 ng/mL~100μg/mL浓度范围,SERS信号强度(ISERS)与CRP浓度的对数值呈良好的线性关系。对人血清样品进行加标试验,回收率为93.1%~109.6%,相对标准偏差(RSD)为2.71%~4.68%(n=3)。该SERS-磁性生物传感器具有制备相对简单、线性范围宽、特异性强、重现性和稳定性好等特点,在临床检测诊断方面具有一定的应用价值。A magnetic capture substrate Fe3O4@PEI@Au@Aptamer-SH was prepared by modifying gold-shell magnetic particles with the sulfhydrylated aptamer of C-reactive protein(CRP);a SERS probe Ag@4-MBA@Aptamer-NH_(2) was prepared by modifying silver nanoparticles with the Raman reporter molecule p-mercaptobenzoic acid(4-MBA)and coupling the aminated aptamer of CRP by amide bond.The magnetic capture substrate,the antigen CRP and the SERS probe formed a sandwich complex,which was magnetically separated and enriched to achieve SERS detection of CRP.The linear relationship between ISERS and CRP concentration was good in the concentration range of 0.1 ng/mL~100μg/mL.The recoveries of human serum samples spiked with SERS ranged from 93.1%~109.6%,and the RSD(n=3)ranged from 2.71%~4.68%.The SERS-magnetic biosensor is relatively simple to prepare,has a wide linear range,good specificity,reproducibility and stability,and has a certain application in clinical detection and diagnosis.

关 键 词：C反应蛋白(CRP) 适配体 SERS 生物传感器 

分 类 号：TP212.3[自动化与计算机技术—检测技术与自动化装置]