Amperometric determination of the activity of protein kinase a using a glassy carbon electrode modified with IgG functionalized gold nanoparticles conjugated to horseradish peroxidase

作者：Zhou, Yunlei; Wang, Minghui; Yin, Huanshun^*; Ai, Shiyun

来源：Microchimica Acta, 2017, 184(9): 3301-3308.

DOI：10.1007/s00604-017-2341-x

• Electrochemical immunosensor
• Phosphoserine antibody
• Enzymatic signal amplification
• Cancer biomarker
• HA-1077 inhibitor
• Targeted drug screening

The authors describe the fabrication of an electrochemical immunosensor for the determination of the activity of protein kinase A (PKA). The method involves (a) electrochemical deposition of gold nanoparticles (AuNPs) on a glassy carbon electrode, (b) PKA-induced catalytic phosphorylation of serine, and (c) the use of phosphoserine antibody and horseradish peroxidase conjugated to IgG on gold nanoparticles (HRP-IgG-AuNPs). The use of AuNPs and HRP-IgG-AuNPs results in large amplification so that the method, at a typical working potential as low as 0.08 V (vs. SCE), has a linear range that extends from 0.1 to 50 activity units per mL, and the detection limit is 0.026 units per mL (at an S/N ratio of 3). The assay is selective (not the least due to a rather low working potential) and well reproducible. It may also be applied to screening for PKA inhibitors and to quantify the PKA activity in human cell lysates.

• 出版日期2017-9
• 单位山东农业大学