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A Rapid and Ultrasensitive Thrombin Biosensor Based on a Rationally Designed Trifunctional Protein
Huayue Zhang1, Lu Yang1, Xiaqing Zhu1
1School of Life Sciences, Tianjin Key Laboratory of Function and Application of Biological Macromolecular Structures, College of Precision Instrument and Opto-Electronics Engineering, Tianjin University, Tianjin, 300072, China.
Advanced Healthcare Materials
|May 15, 2020
Summary
A novel trifunctional protein biosensor enables ultrasensitive and rapid detection of thrombin. This engineered protein achieves the lowest reported limit of detection for thrombin detection in serum, aiding early disease diagnosis.
Area of Science:
- Biotechnology
- Protein Engineering
- Biosensor Development
Background:
- Thrombin detection is crucial for diagnosing and managing thrombin-related diseases.
- Existing biosensors often lack the required sensitivity and speed for early clinical intervention.
Purpose of the Study:
- To develop an ultrasensitive and rapid thrombin biosensor using a rationally designed trifunctional protein.
- To engineer a biosensor with enhanced sensitivity and specificity for thrombin detection.
Main Methods:
- Engineered a trifunctional protein (HTs) incorporating a far-red fluorescent protein (smURFP), hydrophobin (HGFI), and a thrombin cleavage site (TCS).
- Immobilized HTs onto a multiwall plate via HGFI for rapid assay setup.
- Utilized smURFP fluorescence decrease upon thrombin-induced TCS hydrolysis for signal detection.
Main Results:
- Achieved specific recognition and detection of active thrombin in samples.
- Demonstrated a thrombin concentration-dependent decrease in fluorescence intensity.
- Established a limit of detection (LOD) as low as 0.2 am in serum, the lowest reported for any thrombin biosensor.
Conclusions:
- The developed trifunctional protein biosensor offers ultrasensitive and rapid thrombin detection.
- This engineered protein system significantly advances the capabilities of thrombin detection assays.
- Highlights the potential of multifunctional protein engineering for next-generation biosensing applications.
