A ''turn-off'' SERS assay for kinase detection based on arginine N-phosphorylation process

Area of Science:

• Biochemistry
• Analytical Chemistry
• Nanotechnology

Background:

• Protein kinases play crucial roles in cellular signaling pathways.
• Dysregulation of kinases, such as McsB, is implicated in various diseases.
• Sensitive and selective detection methods for kinases are essential for diagnostics.

Purpose of the Study:

• To develop a novel "turn-off" surface-enhanced Raman scattering (SERS) biosensor for the detection of protein arginine kinase McsB.
• To utilize the arginine N-phosphorylation process as the detection mechanism.
• To establish a highly sensitive, selective, and simple assay for kinase detection.

Main Methods:

• Construction of a SERS assay platform using labeled gold nanoparticles (AuNPs).
• Exploitation of the "turn-off" principle based on arginine N-phosphorylation.
• Monitoring changes in SERS intensity due to AuNPs aggregation and disaggregation.

Main Results:

• The assay demonstrated high sensitivity with a detection limit of 46 pM for McsB.
• The biosensor exhibited excellent selectivity, attributed to kinase-specific phosphorylation.
• The method showed improved sensitivity compared to other non-enzymatic amplification techniques.

Conclusions:

• A novel and effective "turn-off" SERS biosensor for McsB detection was successfully established.
• The assay's sensitivity and selectivity make it suitable for biomarker detection in complex samples.
• This platform holds potential for broader applications in detecting other kinases and biomarkers.