北京工业大学 环境科学系,北京 100124
鲁理平,博士,教授,研究方向:环境分析,E-mail:lipinglu@bjut.edu.cn
纸质出版日期:2024-07-15,
收稿日期:2024-04-18,
修回日期:2024-05-06,
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贺彰瑾,杜雨莹,鲁理平.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[J].分析测试学报,2024,43(07):963-970.
HE Zhang-jin,DU Yu-ying,LU Li-ping.Sensitive Detection of Telomerase Activity Based on a G-quadruplex Electrochemiluminescent Biosensor[J].Journal of Instrumental Analysis,2024,43(07):963-970.
贺彰瑾,杜雨莹,鲁理平.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[J].分析测试学报,2024,43(07):963-970. DOI: 10.12452/j.fxcsxb.24041803.
HE Zhang-jin,DU Yu-ying,LU Li-ping.Sensitive Detection of Telomerase Activity Based on a G-quadruplex Electrochemiluminescent Biosensor[J].Journal of Instrumental Analysis,2024,43(07):963-970. DOI: 10.12452/j.fxcsxb.24041803.
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,prognosis,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 luminescent 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 reproducibility. Finally,the sensor was used for the determination of telomerase activity in real human serum and urine samples,and satisfactory results were also obtained.
端粒酶活性G-四链体金属配合物电化学发光生物传感器
telomerase activityG-quadruplexmetal complexelectrochemiluminescencebiosensor
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