基于三脚架DNA探针的电化学发光生物传感器检测外泌体OA北大核心CSTPCD
Tripod DNA Probe-based Electrochemiluminescent Biosensor for Detection of Exosome
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(H2O2)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70～1.4×106 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 the π-π interaction 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(H2O2)electrochemiluminescence(ECL)system.The ECL intensity of the biosensor showed a good linear relationship with logarithm of exosome concentration in the concnetrtion range of 70-1.4×106 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.
贺彰瑾;金鑫;杜雨莹;吴洁宁;鲁理平
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
外泌体三脚架DNA适配体生物传感器电化学发光
ExosomesTripod DNAAptamerBiosensorsElectrochemiluminescence
《分析化学》 2024 (009)
1328-1336 / 9
国家自然科学基金项目(Nos.22276005,21876005)和北京市卓越青年科学家计划项目(No.BJJWZYJH01201910005017)资助. Supported by the National Natural Science Foundation of China(Nos.22276005,21876005)and the Beijing Outstanding Young Scientist Program(No.BJJWZYJH01201910005017).
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