Monitoring casein kinase II at subcellular level via bio-bar-code-based electrochemiluminescence biosensing method
作者： Lifen Wang , Jiajia Song , Xiaofei Wang , Honglan Qi , Qiang Gao
DOI: 10.1016/J.CCLET.2020.06.032
关键词: Chemistry 、 Casein kinase 2 、 Biosensor 、 Detection limit 、 Colloidal gold 、 Peptide 、 Phosphorylated Peptide 、 Biophysics 、 Electrochemiluminescence 、 Ultramicroelectrode
摘要: Abstract A highly sensitive electrochemiluminescence (ECL) biosensing method was developed for monitoring casein kinase II (CK2) at subcellular level via bio-bar-code assay. probe (h-DNA/AuNPs/p-DNA) prepared by conjugating phosphorylated DNA (p-DNA) and hairpin (h-DNA) onto gold nanoparticles (AuNPs) used as a carrier ECL signal reagent (Ru(phen)32+) while specific peptide recognition substance. ultramicroelectrode with diameter of 400 nm fabricated then modified the self-assembly technique to obtain ultramicroelectrode. The on in presence CK2 adenosine 5′-triphosphate, integrated h-DNA/AuNPs/p-DNA through process mediated zirconium cations (Zr4+), finally Ru(phen)32+ intercalated into h-DNA. “signal on” detection range 0.005–0.2 U/mL limit 0.001 U/mL. Additionally, combined efficient phosphorylation vivo bio-bar-code-based method, further applied monitor without tedious fractionation. It found that concentration inserting nucleus higher than cytoplasm HeLa cells. distinct heterogeneity among concentrations single cells observed cellular assessment.
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