College of Chemistry and Material Science, Shandong Agricultural University, Tai'an 271018, PR China.
College of Chemistry and Material Science, Shandong Agricultural University, Tai'an 271018, PR China.
Biosens Bioelectron. 2018 Jun 15;108:89-96. doi: 10.1016/j.bios.2018.02.056. Epub 2018 Feb 27.
Herein, a novel dual-signal amplified photoelectrochemical (PEC) biosensor was successfully developed for the highly selective detection of N-methyladenosine (mA) methylated RNA. The PEC biosensor comprised BiVO-110-TiO heterojunction and gold nanoparticle decorated MoS (MoS-AuNPs) as the photoactive materials, horseradish peroxidase conjugated biotin (HRP-Biotin) as the enzymatic unit, Ag-mediated cytosine pairs (C-Ag-C) as the signal amplification unit, and the anti-mA antibody as the mA methylated RNA recognition unit. Following immunoreaction between mA and the anti-mA antibody, the C-Ag-C structure of the hairpin DNA unfolded, yielding the duplex strand DNA (dsDNA) and releasing Ag ions. Superoxide ions (O) generated by the action of HRP on HO then served as an electron donor, resulting in the deposition of Ag on AuNPs surface and resulting in an increased photocurrent. Based on this change f the photocurrent, mA could be accurately assayed using this dual-signal amplified PEC biosensor. The biosensor showed high selectivity and a very low detection limit of 1.665 pM for mA, and was successfully applied to evaluate the content change of mA in leaves of maize seedling and chicken fetal hepatocytes samples after treatment with chemical mutagen of ethylmethane sulfonate and hormone of insulin, respectively.
本文中,成功开发了一种新型的双信号放大光电化学(PEC)生物传感器,用于高度选择性地检测 N6-甲基腺苷(mA)甲基化 RNA。PEC 生物传感器由 BiVO-110-TiO 异质结和金纳米粒子修饰的 MoS(MoS-AuNPs)作为光活性材料、辣根过氧化物酶偶联生物素(HRP-Biotin)作为酶单元、Ag 介导的胞嘧啶对(C-Ag-C)作为信号放大单元、以及抗 mA 抗体作为 mA 甲基化 RNA 识别单元组成。mA 与抗 mA 抗体发生免疫反应后,发夹 DNA 的 C-Ag-C 结构展开,生成双链 DNA(dsDNA)并释放出 Ag 离子。然后,HRP 在 HO 作用下产生的超氧离子(O)作为电子供体,导致 Ag 在 AuNPs 表面沉积,从而导致光电流增加。基于这种光电流的变化,可以使用这种双信号放大 PEC 生物传感器准确地测定 mA。该生物传感器表现出高选择性和对 mA 的极低检测限为 1.665 pM,并成功应用于分别评估化学诱变剂乙烷磺酸和激素胰岛素处理后玉米幼苗叶片和鸡胎肝细胞样品中 mA 含量的变化。