A highly sensitive ECL biosensor for NF-κB p50 detection based on entropy-driven amplification and CRISPR/Cas12a...

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Biosensor Technology

Background:

• Nuclear factor-kappa B subunit 50 (NF-κB p50) is a key transcription factor implicated in diseases like cancer and inflammation.
• Sensitive detection of NF-κB p50 is critical for clinical diagnostics and monitoring treatment efficacy.

Purpose of the Study:

• To develop a highly sensitive and specific electrochemiluminescence (ECL) biosensor for detecting NF-κB p50.
• To integrate entropy-driven amplification and CRISPR/Cas12a for enhanced signal amplification.

Main Methods:

• Designed an ECL biosensor incorporating entropy-driven amplification and CRISPR/Cas12a.
• Utilized a dsDNA probe and exonuclease III (Exo III) for signal modulation based on NF-κB p50 presence.
• Employed CRISPR/Cas12a system activation for signal enhancement in the absence of the target.

Main Results:

• Achieved a low detection limit of 0.56 pM for NF-κB p50.
• Demonstrated high selectivity and excellent reproducibility of the biosensor.
• Successfully detected NF-κB p50 in complex biological samples like HeLa cell lysates.

Conclusions:

• The developed ECL biosensor offers a sensitive and specific method for NF-κB p50 detection.
• The integration of entropy-driven amplification and CRISPR/Cas12a shows promise for advanced molecular diagnostics.
• This biosensor has significant potential for clinical applications in disease diagnosis and management.