基于G-四链体电化学发光生物传感器灵敏检测端粒酶活性  

Sensitive Detection of Telomerase Activity Based on a G-quadruplex Electrochemiluminescent Biosensor

作　　者：贺彰瑾 杜雨莹 鲁理平[1] HE Zhang-jin;DU Yu-ying;LU Li-ping(Department of Environmental Science,Beijing University of Technology,Beijing 100124,China)

机构地区：[1]北京工业大学环境科学系,北京100124

出　　处：《分析测试学报》2024年第7期963-970,共8页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(22276005,21876005)。

摘　　要：端粒酶活性的分析检测对于癌症诊断、预后治疗等具有重要意义。该研究利用G-四链体(G4) DNA的金属配合物靶向发光探针[Ir(ppy)_(2)(pip)]PF_(6),构建了灵敏检测癌细胞端粒酶活性的电化学发光(ECL)生物传感器。首先将引物DNA组装在电极表面,然后与癌细胞提取的端粒酶共同孵育,具有活性的端粒酶在引物末端生成端粒重复序列从而形成G4结构,随后[Ir(ppy)_(2)(pip)]PF_(6)通过与G4靶向作用结合到延长的序列中。结果表明[Ir(ppy)_(2)(pip)]PF_(6)的ECL信号与端粒酶活性呈现正相关,传感器的电化学发光强度与癌细胞浓度在10-5×10^(5)cell/mL呈现良好的线性关系,检出限(LOD)为3 cell/mL。实验还表明了传感器具有良好的稳定性、适用性、抗干扰性和重现性,最后将该传感器用于测定真实人类血清和尿液样本中的端粒酶活性,得到了满意的结果。The analysis and detection of telomerase activity is important for cancer diagnosis,prog⁃nosis,and treatment.In this study,an electrochemiluminescence(ECL)biosensor was constructed for sensitive detection of telomerase activity in cancer cells using a metal complex-targeted lumines⁃cent probe[Ir(ppy)_(2)(pip)]PF_(6) for G-quadruplex(G4)DNA.Primer DNA was first assembled on the electrode surface and then co-incubated with telomerase extracted from cancer cells.The active telomerase generated telomeric repeats at the end of the primer to form the G4 structure,and then[Ir(ppy)_(2)(pip)]PF_(6) bound to the extended sequence by targeting with G4.The results showed that the ECL signal of[Ir(ppy)_(2)(pip)]PF_(6) showed a positive correlation with telomerase activity,and the ECL intensity of the sensor showed a good linear correlation with the concentration of cancer cells in the range of 10-5×10^(5) cell/mL,with the limit of detection(LOD)of 3 cell/mL.The experiments also demonstrated that the sensor had good stability,applicability,anti-interference ablity,and repro⁃ducibility.Finally,the sensor was used for the determination of telomerase activity in real human se⁃rum and urine samples,and satisfactory results were also obtained.

关 键 词：端粒酶活性 G-四链体 金属配合物 电化学发光 生物传感器 

分 类 号：O646[理学—物理化学] TP212.3[理学—化学]