基于适配体识别的凝血酶均相电化学发光检测方法(英文)  

Homogeneous Electrogenerated Chemiluminescence Aptamer-based Method for the Determination of Thrombin

作　　者：孙丽娟[1] 杨佳[1] 张瑜[1] 漆红兰[1] 高强[1] 张成孝[1] 

机构地区：[1]陕西省生命分析化学重点实验室,陕西师范大学化学与化工学院,陕西西安710062

出　　处：《分析科学学报》2014年第5期651-656,共6页Journal of Analytical Science

基　　金：National Natural Foundation of China(No.21275095,21375084);National Natural Foundation of Shanxi Province(No.2013KJXX-73)

摘　　要：本文以钌联吡啶络合物标记的凝血酶适配体为电化学发光探针,建立了均相电化学发光测定凝血酶的新方法。实验发现待测物凝血酶的存在,使金电极上电化学发光探针的电化学发光强度急剧降低,这是由于电化学发光探针与凝血酶形成了大质量生物复合物,使其扩散系数增大和电化学发光效率降低所致。实验结果表明,电化学发光强度的降低与凝血酶浓度在0.5∽7.5nmol/L范围内呈良好的线性关系。该方法的检测限为0.25nmol/L,对凝血酶测定的相对标准偏差为2.7%（c=5.0nmol/L,n=7）。该方法具有简单灵敏、选择性好和无需探针固定化和冲洗步骤等优点。A novel homogeneous electrogenerated chemiluminescence（ECL）method for the determination of thrombin was developed by employing an ECL probe consisting of thrombin aptamer serving as a recognition element and tris（2,2′-bipyridyl）ruthenium derivatives as an ECL tag.It was found that a strong ECL emission was electrochemically generated from the ECL probe at a gold electrode in the absence of thrombin and decreased markedly in the presence of thrombin.The changes of the integrated ECL intensity was linearly dependent on the concentration of thrombin in the range from 0.5to 7.5nmol/L.The detection limit for thrombin was 0.25nmol/L（S/N=3）and the relative standard derivation for 5.0nmol/L was 2.7%（n=7）.The developed method has advantages such as simplicity,high sensitivity and high selectivity,free of immobilization and washing steps.This work demonstrates that the homogeneous ECL method based on the combination of a high-affinity aptamer with highly sensitive ECL method is a great promising approach for simple and sensitive determination of proteins.

关 键 词：电化学发光 凝血酶 适配体 三(2 2′-联吡啶)钌络合物 

分 类 号：O657.39[理学—分析化学]