基于碳纳米颗粒的荧光适配体传感器用于腺苷脱氨酶的检测  

A novel carbon nanoparticles-based fluorescent aptasensor for detection of adenosine deaminase

作　　者：冯婷婷 王迎超 郭建花 王晓华[3] 杨丽霞 FENG Tingting;WANG Yingchao;GUO Jianhua;WANG Xiaohua;YANG Lixia(Institute of Pharmaceutical and Food Engineering,Shanxi University of Chinese Medicine,Jinzhong 030619;College of Pharmaceutical Sciences,Zhejiang University,Hangzhou 310058;Pharmacy School of Guilin Medical University,Guilin Medical University,Guilin 541004)

机构地区：[1]山西中医药大学中药与食品工程学院,晋中030619 [2]浙江大学药学院,杭州310058 [3]桂林医学院药学院,桂林541004

出　　处：《分析试验室》2020年第8期974-979,共6页Chinese Journal of Analysis Laboratory

基　　金：山西中医药大学博士科研启动基金(2014BK19)项目资助。

摘　　要：设计了一段羧基荧光素(FAM)标记的适配体,该适配体能与腺苷进行高亲和力和强特异性的结合,而不与肌苷发生作用。腺苷脱氨酶可以与腺苷发生脱氨反应,生成肌苷。基于上述原理,构建了碳纳米颗粒-适配体-腺苷荧光适配体传感器用于腺苷脱氨酶的检测。当体系中没有腺苷脱氨酶时,FAM标记的适配体与腺苷紧密结合,而不能被碳纳米颗粒吸附,体系荧光较强;当体系中存在腺苷脱氨酶时,腺苷变成肌苷,肌苷不与适配体结合,此时FAM标记的适配体被碳纳米颗粒吸附,FAM荧光淬灭,体系具有较低的荧光强度。该方法简单、灵敏,线性范围为0.25~3.125 U/mL,检出限为0.18 U/mL。与其它蛋白分子相比,方法对腺苷脱氨酶的检测具有高特异性。构建的传感器简单,再生性好,可用于标准加入法检测小鼠血清中的腺苷脱氨酶。The carboxyfluorescein(FAM)labeled aptamer that had a strong affinity for adenosine,was synthesized first.Then,the mixing of FAM-labeled aptamer with carbon nanoparticles(CNPs)formed a selfassembly of two components via the electrostatic interactions between them.As a result,the fluorescence of the FAM-labeled aptamer was quenched by the efficient electron transfer for detection of adenosine deaminase.In the presence of adenosine,the FAM-labeled aptamer in the self-assembly would be released from the CNPs surface,thereby causing the fluorescence recovery.Upon reaction with adenosine deaminase(ADA),the fluorescence of the complex was greatly quenched as a consequence of the removal of adenosine converted into inosine that has no affinity for this aptamer.This behaviour has led to the development of a rapid and sensitive fluorescent method for assaying ADA activity,with a detection limit of 0.18 U/m L.This approach is simplely compared to the existing ones since the CNPs based sensor is easy to be assembled and the detection can be achieved without the involvement of complicated procedures.Furthermore,the applicability of the method has been demonstrated by detecting ADA in mouse serum samples.

关 键 词：碳纳米颗粒 适配体 荧光适配体传感器 腺苷 腺苷脱氨酶 

分 类 号：O657.39[理学—分析化学]