Ultrasensitive Chemiluminescence Probes Designed from Covalent Inhibitors for SRAS-CoV-2 M^pro Detection

Area of Science:

• Biochemistry
• Molecular Biology
• Infectious Diseases

Background:

• Long COVID presents ongoing health challenges, necessitating improved methods for SARS-CoV-2 monitoring.
• The SARS-CoV-2 main protease (Mpro) is a crucial viral target and a potential biomarker due to its conserved nature and specificity.

Purpose of the Study:

• To develop efficient chemiluminescence probes for detecting SARS-CoV-2 Mpro.
• To create a tool for rapid drug screening and monitoring of viral activity.

Main Methods:

• Design and synthesis of chemiluminescence probes based on Mpro-targeted covalent inhibitors.
• Evaluation of probe performance including response time, limit of detection (LoD), selectivity, and stability.
• Validation of probe mechanism and application in cell-based imaging of authentic SARS-CoV-2 infected cells.

Main Results:

• Developed several Mpro-targeted chemiluminescence probes.
• Identified MPCL-2 as the optimal probe, demonstrating a rapid response (<20 min) and a very low LoD (0.11 nM).
• MPCL-2 showed high selectivity, stability, and was successfully used for real-time, in-situ imaging in infected cells, with potential for in vivo applications.

Conclusions:

• MPCL-2 offers superior performance for SARS-CoV-2 Mpro detection compared to existing methods.
• The probe is applicable for drug screening and monitoring viral activity.
• The substrate design strategy can be adapted for developing probes against other pathogens.