Real-time in vitro detection of cellular H2O2 under camptothecin stress using horseradish peroxidase, ionic liquid, and carbon nanotube-modified carbon fiber ultramicroelectrode

SENSORS AND ACTUATORS B-CHEMICAL

卷:245

Article

Ren, Qiong-Qiong¹  Wu, Jing²  Zhang, Wen-Chao¹  Wang, Chang¹  Qin, Xin¹  Liu, Guo-Chang³  Li, Zhen-Xin¹  Yu, Yi¹ 

[1] Xinxiang Med Univ, Sch Biomed Engn, Xinxiang, Peoples R China

[2] Xinxiang Med Univ, Inst Lung & Mol Therapy, Xinxiang, Peoples R China

[3] Huazhong Univ Sci & Technol, Britton Chance Ctr Biomed Photon, Wuhan Natl Lab Optoelect,Coll Life Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab,Dept Biom, Wuhan 430074, Peoples R China

关键词: Hydrogen peroxide;    Single-walled carbon nanotubes;    Horseradish peroxidase;    Room-temperature ionic liquids;    Carbon fiber;    Real-time monitoring;   

DOI  :  10.1016/j.snb.2017.02.001

来源: Elsevier

【 摘 要 】

Single-walled carbon nanotubes (SWCNTs), horseradish peroxidase (HRP), and 1-butyl-3methylimidazolium tetrafluoroborate (BMIM center dot BF4) were employed to construct a cellular H2O2 sensor based on direct electron transfer. At a working potential of 0.35V, HRP-BMIM center dot BF4/SWCNTs/CFUME showed a dynamic range of up to 10.2 mu M with a low detection limit of 0.13 mu M (S/N =3) and a high sensitivity of 4.25 A/M cm(2). The apparent Michaelis-Menten constant (K-m,K-app) was estimated to be as low as 15.4 mu M, which suggested that HRP molecules entrapped in the BMIM center dot BF4 and SWCNTs immobilized at the carbon fiber ultramicroelectrode maintained a very high affinity. Because of the extremely small dimension and low working potential, HRP-BMIM center dot B-4/SWCNTs/CFUME enabled direct amperometric real-time monitoring of H2O2 in HeLa cells treated with the anticancer drug, camptothecin, without requiring complex data processing and extra surface coatings to prevent interference. HRP BMIM center dot BF4/SWCNTs/CFUME testing clearly showed that the H2O2 level significantly increased in HeLa cells under camptothecin stress. When HeLa cells were cultured in medium without camptothecin, the H2O2 level remained stable during the whole measurement process. These results indicate that HRP-BMIM center dot BF4/SWCNTs/CFUME could be a powerful tool for real-time investigation of cellular H2O2 level, especially under anticancer drug stress. This method could provide in-depth insights regarding the occurrence, development, and apoptosis of tumors. (C) 2017 The Authors. Published by Elsevier B.V.

【 授权许可】

Free