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基于具有高光致发光效率的自增强电致化学发光钌配合物掺杂的金属有机骨架的端粒酶活性超高灵敏检测。

Ultrasensitive Assay for Telomerase Activity via Self-Enhanced Electrochemiluminescent Ruthenium Complex Doped Metal-Organic Frameworks with High Emission Efficiency.

机构信息

Key Laboratory of Luminescence and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University , Chongqing 400715, People's Republic of China.

Department of Clinical Biochemistry, Laboratory Sciences, Third Military Medical University , 30 Gaotanyan Street, Shapingba District, Chongqing 400038, China.

出版信息

Anal Chem. 2017 Mar 7;89(5):3222-3227. doi: 10.1021/acs.analchem.7b00259. Epub 2017 Feb 23.

DOI:10.1021/acs.analchem.7b00259
PMID:28194971
Abstract

Here, an ultrasensitive "off-on" electrochemiluminescence (ECL) biosensor was proposed for the determination of telomerase activity by using a self-enhanced ruthenium polyethylenimine (Ru-PEI) complex doped zeolitic imidazolate framework-8 (Ru-PEI@ZIF-8) with high ECL efficiency as an ECL indicator and an enzyme-assisted DNA cycle amplification strategy. The Ru-PEI@ZIF-8 nanocomposites were synthesized by self-enhanced Ru-PEI complex doping during the growth of zeolitic imidazolate framework-8 (ZIF-8), which presented high ECL efficiency and excellent stability. Furthermore, owing to the porosity of Ru-PEI@ZIF-8, the self-enhanced Ru-PEI complex in the outer layer and inner layer of self-enhanced Ru-PEI@ZIF-8 could be excited by electrons causing the utilization ratio of the self-enhanced ECL materials to be remarkably increased. To further improve the sensitivity of the proposed biosensor, the telomerase activity signal was converted into the trigger DNA signal which was further amplified by an enzyme-assisted DNA recycle-amplification strategy. The proposed ECL biosensor presented great performance for telomerase activity detection from 5 × 10 to 10 Hela cells with a detection limit of 11 cells. Moreover, this method was applied in the detection of telomerase activity from cancer cells treated with an anticancer drug, which indicated the proposed method held potential application value as an evaluation tool in anticancer drug screening.

摘要

这里,提出了一种超灵敏的“开-关”电化学发光(ECL)生物传感器,用于通过使用具有高 ECL 效率的自增强钌聚乙烯亚胺(Ru-PEI)配合物掺杂沸石咪唑骨架-8(Ru-PEI@ZIF-8)作为 ECL 指示剂和酶辅助 DNA 循环扩增策略来测定端粒酶活性。Ru-PEI@ZIF-8 纳米复合材料通过沸石咪唑骨架-8(ZIF-8)生长过程中的自增强 Ru-PEI 配合物掺杂合成,具有高 ECL 效率和优异的稳定性。此外,由于 Ru-PEI@ZIF-8 的多孔性,外层和内层的自增强 Ru-PEI 配合物可以被电子激发,从而显著提高自增强 ECL 材料的利用率。为了进一步提高所提出的生物传感器的灵敏度,将端粒酶活性信号转换为触发 DNA 信号,然后通过酶辅助 DNA 循环扩增策略进一步放大。该 ECL 生物传感器在检测 5×10 至 10 个 Hela 细胞的端粒酶活性方面表现出优异的性能,检测限为 11 个细胞。此外,该方法还应用于抗癌药物处理的癌细胞中端粒酶活性的检测,表明该方法作为抗癌药物筛选的评估工具具有潜在的应用价值。

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