石墨烯增敏的SPR技术应用于HIV1 p24抗原的检测  

Graphene Assisted detection of HIV1 p24 antigen by Surface Plasmon Resonance Technique

作　　者：彭芳 张良 何建安 PENG Fang;ZHANG Liang;HE Jianan(Guangzhou center for disease control and prevention,Guangdong Guangzhou 510500;Shenzhen Academy of Inspection and Quarantine,Guangdong Shenzhen 518010)

机构地区：[1]广州市荔湾区疾病预防控制中心,广东广州510500 [2]深圳市检验检疫科学研究院,广东深圳518045

出　　处：《生物化工》2021年第3期4-8,共5页Biological Chemical Engineering

基　　金：广州市卫生健康科技项目(20181A011107)。

摘　　要：目的:建立石墨烯增敏的表面等离子体共振技术(Surface Plasmon Resonance,SPR)用于检测HIV1 p24,为HIV感染者的早期快速筛查提供一种检测方法。方法:石墨烯通过静电连接在采用聚乙二醇高分子处理的SPR芯片表面,HIV1 p24特异性抗体通过石墨烯表面的羧基官能团和HIV1 p24抗体表面的氨基官能团共价结合作用固定在芯片的表面,建立一种石墨烯增敏的无标记、快速的表面等离子共振技术用于检测HIV1 p24抗原,通过优化芯片分析条件,检测HIV1 p24抗原,分析石墨烯增敏的SPR芯片技术的检测性能。结果:石墨烯增敏的SPR芯片技术检测HIV1 p24抗原的重复性、敏感性与特异性均较好;检测灵敏度为10 ng/mL,相较于相同条件传统Au-SPR生物传感器检测得到的最低检测限(63 ng/mL),降低了至少6倍。结论:初步构建的石墨烯增敏的SPR芯片技术,有望为HIV感染者的早期筛查提供一个有力的技术平台。Objective:A surface plasmon resonance(SPR)technique based on grapheme oxide(GO)for determination of HIV1 p24 so as to provide a simple and rapid method for early screening of HIV;Methods:GO was conjugated onto the surface of the SPR chip surface via electrostatic interaction after the chip surface was activated with cysteamine.Subsequently,HIV1 p24 antibody was covalently immobilized on the chip surface through binding between carboxylic groups on the GO surface and amine groups on the protein surface.Finally,label-free and rapid detection of HIV1 p24 was carried out by the SPR biosensor via protein–protein interaction.The analysis conditions of GO based SPR chip were optimized and then the HIV1 p24 antigens were detected;Results:The repeatability,sensitivity and specificity of graphene based SPR chip technique for detecting HIV1 p24 antigen all were better.Under the optimized conditions,limits of detection(LODs)was 10ng/ml for P24 antigen in a buffer solution.Conclusion These data suggest that GO based SPR chip technique can be used to detect HIV1 p24 antigen,which would be potentially useful for HIV early screening in both endemic and non-endemic areas.

关 键 词：石墨烯 SPR技术 HIV1 P24抗原 早期筛查 

分 类 号：TP212.3[自动化与计算机技术—检测技术与自动化装置]