A highly sensitive electrochemiluminescence assay for protein kinase based on double-quenching of graphene quantum dots by G-quadruplex-hemin and gold nanoparticles

Biosensors & Bioelectronics
Jinquan LiuLi Wen

Abstract

A highly sensitive electrochemiluminescence (ECL) strategy was developed for the protein kinase A (PKA) activity and inhibition assay based on double-quenching of graphene quantum dots (GQDs) ECL by G-quadruplex-hemin DNAzyme and gold nanoparticles (AuNPs). In this strategy, the GQDs were modified onto the indium-tin oxide (ITO) electrode and further assembled with substrate peptide of target protein kinase through covalent coupling, which can exhibit high and stable ECL signal. The AuNPs, functionalized with the phosphorylated DNA and G-quadruplex-hemin DNAzyme via Au-S chemistry, were selected as quenching probes. In the presence of PKA, the peptide on the electrode was phosphorylated and the AuNPs functionalized with the phosphorylated DNA and G-quadruplex-hemin DNAzyme were subsequently integrated onto the phosphorylated peptide by Zr(4+). Owing to the reduction of coreactant H2O2 resulting from G-quadruplex-hemin DNAzyme catalytic reaction and the ECL energy transfer (ECL-RET) between AuNPs and GQDs, the ECL intensity of GQDs was significantly decreased. By taking advantage of the double-quenching effect, this assay can detect PKA with a linear range of 0.05 to 5UmL(-1) and a detection limit of 0.04UmL(-1). In addition, th...Continue Reading

Citations

Jul 15, 2015·Biochemical and Biophysical Research Communications·Yuri Volkov
Apr 12, 2018·International Journal of Molecular Sciences·Nawal Ahmad AlarfajHesham Farouk Oraby
Feb 13, 2016·Chembiochem : a European Journal of Chemical Biology·Chun-Chi WangHuan-Tsung Chang
Mar 25, 2019·International Journal of Molecular Sciences·Zhiyong YanYang Liu
Nov 9, 2017·Chemical Society Reviews·Yuanhong XuShaojun Guo

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