Department of Medical IT Convergence, Kumoh National Institute of Technology, Gumi 39177, Korea.
Korea Institute of Toxicology, Jeongeup 56212, Korea.
Biosensors (Basel). 2022 Feb 10;12(2):107. doi: 10.3390/bios12020107.
Nicotinamide adenine dinucleotide (NADH) is an important cofactor involved in metabolic redox reactions in living cells. The detection of NADH in living animal cells is a challenge. We developed a one-step monitoring method for NADH via an electrocatalytic reaction that uses a surface-modified, screen-printed electrode (SPE) having a redox active monolayer 4'-mercapto-N-phenlyquinone diamine (NPQD) formed by a self-assembled monolayer (SAM) of an aromatic thiol, 4-aminothiophenol (4-ATP). This electrode has a limit of detection (LOD) of 0.49 μM and a sensitivity of 0.0076 ± 0.0006 μM/μA in cell culture media, which indicates that it retains its selectivity. The applicability of this NADH sensor was demonstrated for the first time by cell viability monitoring via NADH-sensing in cell culture supernatants.
烟酰胺腺嘌呤二核苷酸 (NADH) 是活细胞内代谢氧化还原反应中的重要辅酶。在活的动物细胞中检测 NADH 是一项挑战。我们开发了一种通过电催化反应一步监测 NADH 的方法,该反应使用表面修饰的、带有氧化还原活性单层的丝网印刷电极 (SPE),该单层由芳基硫醇 4-氨基苯硫酚 (4-ATP) 的自组装单层 (SAM) 形成 4'-巯基-N-苯醌二胺 (NPQD)。该电极在细胞培养液中的检测限 (LOD) 为 0.49 μM,灵敏度为 0.0076 ± 0.0006 μM/μA,表明其保持选择性。首次通过细胞培养上清液中 NADH 检测的细胞活力监测,证明了这种 NADH 传感器的适用性。