Zhou Sai-Sai, Zhang Lin, Cai Qi-Yong, Dong Zhen-Zhen, Geng Xin, Ge Jia, Li Zhao-Hui
College of Chemistry and Molecular Engineering, Zhengzhou University, No. 100 Science Road, Zhengzhou, Henan, 450001, China.
Anal Bioanal Chem. 2016 Sep;408(24):6711-7. doi: 10.1007/s00216-016-9788-1. Epub 2016 Jul 25.
A label-free fluorescence assay has been developed for sensitive and selective detection of adenosine triphosphate (ATP) by using poly(thymine) (poly T)-templated copper nanoparticles (CuNPs) as fluorescent indicator. In our design, ATP aptamer was split into two fragments, both of which were elongated with poly T strands that can be utilized as efficient template for the formation of copper nanoparticles through the reduction of copper ions by sodium ascorbate. In the presence of ATP, the two split aptamers could be dragged to form aptamer-ATP aptamer complex, which drew the poly T strands close to each other and induced a remarkable fluorescence enhancement of poly T-templated CuNPs. Thus, an elevated fluorescence enhancement of poly T-templated CuNPs was obtained with the increase in ATP concentration. Under optimized conditions, a good linear range for ATP detection was realized from 100 nM to 100 μM with a detection limit of 10.29 nM. In addition, the application of this biosensing system in complex biological matrix was demonstrated with satisfactory results. This assay provided a simple, label-free, cost-effective, and sensitive platform for the detection of ATP.
通过使用聚胸腺嘧啶(poly T)模板化的铜纳米颗粒(CuNPs)作为荧光指示剂,开发了一种用于灵敏且选择性检测三磷酸腺苷(ATP)的无标记荧光检测方法。在我们的设计中,ATP适配体被拆分成两个片段,这两个片段都用poly T链进行了延伸,通过抗坏血酸钠还原铜离子,poly T链可作为形成铜纳米颗粒的有效模板。在ATP存在的情况下,两个拆分的适配体可被拉到一起形成适配体-ATP适配体复合物,这使得poly T链彼此靠近,并导致poly T模板化的CuNPs荧光显著增强。因此,随着ATP浓度的增加,poly T模板化的CuNPs荧光增强升高。在优化条件下,实现了ATP检测的良好线性范围为100 nM至100 μM,检测限为10.29 nM。此外,该生物传感系统在复杂生物基质中的应用也得到了令人满意的结果。该检测方法为ATP检测提供了一个简单、无标记、经济高效且灵敏的平台。