Qiang Weibing, Hu Hongting, Sun Liang, Li Hui, Xu Danke
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , 22 Hankou Road, Nanjing, Jiangsu 210093, China.
Anal Chem. 2015 Dec 15;87(24):12190-6. doi: 10.1021/acs.analchem.5b03075. Epub 2015 Nov 20.
A nanocomplex was developed for molecular sensing in living cells, based on the fluorophore-labeled aptamer and the polydopamine nanospheres (PDANS). Due to the interaction between ssDNA and PDANS, the aptamer was adsorbed onto the surface of PDANS forming the aptamer/PDANS nanocomplex, and the fluorescence was quenched by PDANS through Förster resonance energy transfer (FRET). In vitro assay, the introduction of adenosine triphosphate (ATP) led to the dissociation of the aptamer from the PDANS and the recovery of the fluorescence. The retained fluorescence of the nanocomplex was found to be linear with the concentration of ATP in the range of 0.01-2 mM, and the nanocomplex was highly selective toward ATP. For the strong protecting capability to nucleic acids from enzymatic cleavage and the excellent biocompatibility of PDANS, the nanocomplex was transported into cells and successfully realized "signal on" sensing of ATP in living cells; moreover, the nanocomplex could be employed for ATP semiquantification. This design provides a strategy to develop biosensors based on the polydopamine nanomaterials for intracellular molecules analysis. For the advantages of polydopamine, it would be an excellent candidate for many biological applications, such as gene and drug delivery, intracellular imaging, and in vivo monitoring.
基于荧光团标记的适配体和聚多巴胺纳米球(PDANS),开发了一种用于活细胞分子传感的纳米复合物。由于单链DNA与PDANS之间的相互作用,适配体吸附在PDANS表面形成适配体/PDANS纳米复合物,并且通过荧光共振能量转移(FRET),荧光被PDANS淬灭。在体外分析中,引入三磷酸腺苷(ATP)导致适配体从PDANS上解离并使荧光恢复。发现纳米复合物保留的荧光与ATP浓度在0.01 - 2 mM范围内呈线性关系,并且该纳米复合物对ATP具有高度选择性。由于PDANS对核酸具有强大的酶切保护能力和出色的生物相容性,纳米复合物被转运到细胞中,并成功实现了活细胞中ATP的“开启”传感;此外,该纳米复合物可用于ATP的半定量分析。这种设计提供了一种基于聚多巴胺纳米材料开发用于细胞内分子分析的生物传感器的策略。鉴于聚多巴胺的优势,它将是许多生物应用的优秀候选者,如基因和药物递送、细胞内成像以及体内监测。