Hu Ping, Zhang Ying, Wang Dandan, Qi Guohua, Jin Yongdong
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China.
University of Science and Technology of China, Hefei, Anhui 230026, China.
Anal Chem. 2021 Mar 9;93(9):4240-4245. doi: 10.1021/acs.analchem.0c05004. Epub 2021 Feb 24.
Glutathione (GSH) level in cells maintains redox homeostasis, so the detection of intracellular GSH is crucial for many pathological processes. In the present study, we developed a cystamine-functionalized glass nanopore (cG-nanopore) for the specific detection of GSH in single cells through the Michael addition reaction between the amino group of cystamine and polydopamine adhered to the inner wall of the glass nanopore. The cellular GSH will cleave the disulfide bond of cystamine and induce a negative charge increase on the residual surface of the modified nanopore, thus producing a sensitive response on the current-voltage curves. The developed sensing platform showed a good response relationship with the GSH concentration and an excellent selectivity against interfering substances in cells. The variation of GSH content in single HeLa and H8 cells incubated with DOX was then further quantified by this method.
细胞内的谷胱甘肽(GSH)水平维持着氧化还原稳态,因此检测细胞内GSH对于许多病理过程至关重要。在本研究中,我们开发了一种胱胺功能化玻璃纳米孔(cG-纳米孔),通过胱胺的氨基与附着在玻璃纳米孔内壁的聚多巴胺之间的迈克尔加成反应,用于特异性检测单细胞中的GSH。细胞内的GSH会裂解胱胺的二硫键,并导致修饰纳米孔残余表面的负电荷增加,从而在电流-电压曲线上产生灵敏响应。所开发的传感平台与GSH浓度呈现出良好的响应关系,并且对细胞内干扰物质具有出色的选择性。然后通过该方法进一步定量了用阿霉素处理的单个HeLa细胞和H8细胞中GSH含量的变化。