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Nanosensors to Detect Protease Activity In Vivo for Noninvasive Diagnostics
Published on: July 16, 2018
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A highly sensitive NanoLuc-based protease biosensor for detecting apoptosis and SARS-CoV-2 infection
Masashi Arakawa1,2, Akiho Yoshida3,4, Shinya Okamura3,4
1Department of Biochemistry and Molecular Biology, Faculty of Agriculture and Life Science, Hirosaki University, 3 Bunkyo-Cho, Hirosaki-Shi, Aomori, 036-8561, Japan.
Scientific Reports
|January 31, 2023
Summary
A novel protease biosensor, FlipNanoLuc, offers highly sensitive detection of protease activity. This breakthrough enables better monitoring of apoptosis and viral infections like SARS-CoV-2.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Proteases are crucial in biological processes like apoptosis and viral infections.
- Existing protease biosensors struggle to reliably detect low endogenous protease activity.
Purpose of the Study:
- To develop a highly sensitive protease biosensor system.
- To enhance detection of apoptosis and viral infections using the novel biosensor.
Main Methods:
- Development of FlipNanoLuc biosensor using Oplophorus gracilirostris NanoLuc luciferase.
- Incorporation of NanoBiT technology and CL1-PEST1 degradation tag for signal amplification.
- Application of the biosensor for detecting caspase activity in apoptosis and coronavirus 3CL-protease activity.
Main Results:
- FlipNanoLuc achieved over 500-fold luminescence increase in protease-overexpressing cells.
- Caspase sensor detected staurosporine-induced apoptosis with a sixfold luminescence increase.
- Coronavirus 3CL-protease sensor detected OC43 and SARS-CoV-2 with tenfold and 20-fold luminescence increases, respectively.
Conclusions:
- The FlipNanoLuc system provides a highly sensitive platform for protease detection.
- This biosensor is effective for monitoring apoptosis and viral infections, including SARS-CoV-2.
- Further applications in studying protease-related biological processes are promising.
