基于金纳米颗粒的裂分型适配体传感器检测三磷酸腺苷  

A Split-Aptamer Sensor for Detection of Adenosine Triphosphate Based on Gold Nanoparticles

作　　者：任林娇 彭政 孟晓龙 张培 秦自瑞 徐晓萍 徐鹏 姜利英 REN Lin-Jiao;PENG Zheng;MENG Xiao-Long;ZHANG Pei;QIN Zi-Rui;XU Xiao-Ping;XU Peng;JIANG Li-Ying(College of Electrical and Information,Zhengzhou University of Light Industry Engineering,Zhengzhou 450000,China)

机构地区：[1]郑州轻工业大学电气信息工程学院,郑州450000

出　　处：《分析化学》2022年第3期405-412,I0001,I0002,共10页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.62073299,61801436);河南省高校科技创新团队项目(No.20IRTSTHN017);河南省重点研发与推广项目(No.202102210186);河南省高等学校重点科研项目(No.21A410001)资助。

摘　　要：构建了一种基于金纳米颗粒(Gold nanoparticles,AuNPs)的裂分型适配体荧光传感器,用于检测三磷酸腺苷(ATP)。将ATP核酸适配体序列分裂为P1和P2两个片段,在P1的5′端修饰荧光基团羧基荧光素(FAM),3′端巯基化修饰的P2通过Au—S键以自组装的方式修饰在AuNPs表面。未加入ATP时,P1与P2不结合,此时P1远离AuNPs,荧光信号较强;加入ATP后,可形成P1-ATP-P2的三明治结构,P15′端的FAM荧光基团靠近AuNPs,发生荧光共振能量转移,导致荧光信号减弱。对AuNPs与P2的结合、浓度比例以及加入P1和ATP后的稳定时间等实验条件进行了优化。在最优的条件下,传感器的荧光强度与ATP浓度在0.03~3.33 nmol/L和3.33~15 nmol/L浓度范围内呈两段的线性关系,检出限为0.03 nmol/L。该传感器对ATP表现出良好的特异性。A split aptamer sensor based on gold nanoparticles(AuNPs)was constructed for detection of adenosine triphosphate(ATP).The ATP aptamer was splited into two fragments(P1 and P2),of which the 5′end of P1 fragment was modified by carboxyfluorescein(FAM)fluorophore,while the P2 fragment with 3′end thiol-functionalization was modified on the AuNPs surface through the self-assembly method by Au-S bond.It was found that the P1 and P2 fragments could not combine together before adding ATP,and the distance between P1 fragments and AuNPs was relatively far,which resulted in strong fluorescence signal.However,after adding ATP into the sensing system,the sandwich structure of P1-ATP-P2 formed,and the FAM fluorophore in the 5′end of P1 fragments approached to the AuNPs,leading to the decrease of the fluorescence intensity due to fluorescence resonance energy transfer effect.Under optimal experimental conditions,the fluorescence intensity of the sensor showed linear relationship with ATP concentration in the range of 0.03-3.33 nmol/L and 3.33-15 nmol/L,respectively,with a detection limit(S/N=3)of 0.03 nmol/L.Furthermore,the constructed sensor had excellent specificity for detection of ATP.

关 键 词：裂分型核酸适配体 荧光传感器 金纳米颗粒 荧光共振能量转移 三磷酸腺苷 

分 类 号：O657.3[理学—分析化学] Q503[理学—化学]