荧光过氧化氢酶纳米传感器的制备及性能  

Ratiometric Fluorescent Enzymatic Nanosensors for Intracellular Hydrogen Peroxide

作　　者：李春霞 聂方 聂雅坤 王小卉 彭洪尚[2] NIE Fang;NIE Ya-kun;WANG Xiao-hui;PENG Hong-shang(College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China;College of Science, Minzu University of China, Beijing 100081, China;School of Electronic Engineering, Beijing University of Posts and Telecommunications, Beijing 100876, China)

机构地区：[1]中央民族大学生命与环境科学学院,北京100081 [2]中央民族大学理学院,北京100081 [3]北京邮电大学电子工程学院,北京100876

出　　处：《发光学报》2020年第7期826-833,共8页Chinese Journal of Luminescence

基　　金：国家自然科学基金(61775245);中央民族大学“111计划”资助项目。

摘　　要：过氧化氢(H2O2)是活性氧类的主要标志物,它与多种疾病如神经退行性疾病密切相关。本文设计了一种检测细胞内过氧化氢的荧光过氧化氢酶纳米传感器。这种纳米传感器由含多聚赖氨酸、辣根过氧化物酶的生物相容性外壳和含有氧探针的多孔聚合物基质的氧传感核组成。辣根过氧化物酶(HRP)催化H2O2生成氧,进而通过荧光氧气探针进行探测。该酶纳米传感器的流体动力学尺寸约为270 nm,zeta电位为-18 mV,具有良好的生物相容性。它的荧光比率和时间分辨荧光均对H2O2高度敏感。此外,该纳米传感器可以被活细胞有效地摄取,从而可以用TRF方式灵敏地检测细胞内的H2O2浓度。结果表明,本实验制备的酶纳米传感器有望进一步用于监测H2O2相关的细胞生化反应,如氧化应激。Hydrogen peroxide(H2O2)is the main marker of reactive oxygen species and closely related to various diseases such as neurodegenerative diseases.In this work,we presented an enzymatic nanosensor for fluorescence sensing of intracellular hydrogen peroxide.The nanosensor consisted of a biocompatible shell of poly-l-lysine and horseradish peroxidase(HRP),and an oxygen sensing core from a porous polymeric matrix containing oxygen probes.H2O2 was catalyzed by HRP to yield the product of oxygen,which was reported by the sensing core in the form of quenched fluorescence.The enzymatic nanosensors had a hydrodynamic size of about 270 nm,a zeta potential of-18 mV and good biocompatibility.Their fluorescence were highly sensitive to H2O2 in both ratiometric and the time-resolved fluorescence.Moreover,the nanosensors can be efficiently internalized by live cells,thereby intracelluar H2O2 was sensitively detected with TRF modality.These results suggest that our enzymatic nanosensors may be used for monitoring H2O2-related cellular events,such as oxidative stress.

关 键 词：过氧化氢 纳米传感器 时间分辨荧光 比率荧光 氧化应激 

分 类 号：TP212[自动化与计算机技术—检测技术与自动化装置]