Protein phosphorylation detection using dual-mode field-effect devices and nanoplasmonic sensors.
Phosphorylation by kinases is an important post-translational modification of proteins. It is a critical control for the regulation of vital cellular activities, and its dysregulation is implicated in several diseases. A common drug discovery approach involves, therefore, time-consuming screenings of large libraries of candidate compounds to identify novel inhibitors of protein kinases. In this work, we propose a novel method that combines localized surface plasmon resonance (LSPR) and electrolyte insulator semiconductor (EIS)-based proton detection for the rapid identification of novel protein kinase inhibitors. In particular, the selective detection of thiophosphorylated proteins by LSPR is achieved by changing their resonance properties via a pre-binding with gold nanoparticles. In parallel, the EIS field-effect structure allows the real-time electrochemical monitoring of the protein phosphorylation by detecting the release of protons associated with the kinases activity. This innovative combination of both field-effect and nanoplasmonic sensing makes the detection of protein phosphorylation more reliable and effective. As a result, the screening of protein kinase inhibitors becomes more rapid, sensitive, robust and cost-effective.
Department of Electronic &Electrical Engineering, University of Bath, Bath BA2 7AY, United Kingdom.
Nikhil
N
Bhalla
Department of Chemical Engineering, University of Bath, Bath BA2 7AY, United Kingdom.
Mirella
M
Di Lorenzo
Department of Pharmacy &Pharmacology, University of Bath, Bath BA2 7AY, United Kingdom.
Giordano
G
Pula
Department of Electronic &Electrical Engineering, University of Bath, Bath BA2 7AY, United Kingdom.
Pedro
P
Estrela
D015374:Biosensing Techniques; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*
D011506:Proteins