基于三脚架DNA探针的电化学发光生物传感器检测外泌体  

Tripod DNA Probe-based Electrochemiluminescent Biosensor for Detection of Exosome

作　　者：贺彰瑾 金鑫[1] 杜雨莹 吴洁宁 鲁理平[1] HE Zhang-Jin;JIN Xin;DU Yu-Ying;WU Jie-Ning;LU Li-Ping(Key Laboratory of Beijing on Regional Air Pollution Control,Faculty ofEnvironment and Life,Beijing University of Technology,Beijing 100124,China)

机构地区：[1]北京工业大学环境科学系,区域大气复合污染防治北京市重点实验室,北京100124

出　　处：《分析化学》2024年第9期1328-1336,共9页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.22276005,21876005);北京市卓越青年科学家计划项目(No.BJJWZYJH01201910005017)资助。

摘　　要：外泌体(Exosome)的分析检测对于疾病的早期诊断和治疗具有重要意义。本研究利用碱基与石墨烯的π-π相互作用将三螺旋结构DNA(tsDNA)组装在电化学还原氧化石墨烯(ERGO)的表面,制备了三脚架(Tripod)DNA探针,并通过悬于探针顶端的膜蛋白(CD63)适配体(apt)序列特异性捕获外泌体,同时引入血红素增强鲁米诺(Luminol)-过氧化氢(H_(2)O_(2))电化学发光(ECL)体系的信号强度。此生物传感器的ECL强度与外泌体浓度在70~1.4×10^(6)particles/μL范围内呈良好的线性关系,检出限低至30 particles/μL(S/N=3)。这种三脚架结构的DNA较双链或单链DNA具有大的探针间距,使其更有利于较大目标物的捕获和检测,有望应用于临床样品的分析测定。The detection of exosomes is of great significance for early diagnosis and treatment of cancer.In this study,based on theπ-πinteraction of the adenine bases of DNA with graphene,triple-stranded DNA(tsDNA)was assembled on the surface of electrochemically reduced graphene oxide(ERGO)to obtain a tripod DNA probe.The exosomes was specifically captured by exosome membrane protein(CD63)aptamer(apt)sequence at the top of tsDNA,and the introduction of hemin effectively enhanced the signal intensity of the Luminol-hydrogen peroxide(H_(2)O_(2))electrochemiluminescence(ECL)system.The ECL intensity of the biosensor showed a good linear relationship with logarithm of exosome concentration in the concnetrtion range of 70–1.4×10^(6)particles/μL,and the detection limit was 30 particles/μL(S/N=3).This tripod-structured DNA probe facilitated the capture and detection of larger targets,and was expected to be applied to the analysis of actual clinical samples.

关 键 词：外泌体 三脚架DNA 适配体 生物传感器 电化学发光 

分 类 号：O657.3[理学—分析化学] TP212.3[理学—化学] R446[自动化与计算机技术—检测技术与自动化装置]