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A simple fluorescence assay for trypsin through a protamine-induced carbon quantum dot-quenching aggregation platform
Yiping Chen1, Zuan Lin2, Chenfang Miao2
1Department of Interventional Radiology, The First Affiliated Hospital of Fujian Medical University Fuzhou 350005 China.
RSC Advances
|May 6, 2022
Summary
A new "turn-on" fluorescence method detects trypsin (Try) using carbon quantum dots (CQDs) and protamine (Pro). This simple assay is valuable for disease diagnosis and drug screening.
Area of Science:
- Biochemistry
- Nanotechnology
- Analytical Chemistry
Background:
- Trypsin (Try) plays a crucial role in various physiological and pathological processes, making its accurate detection vital for disease diagnosis.
- Existing detection methods for trypsin often lack simplicity, sensitivity, or specificity.
- Carbon quantum dots (CQDs) offer unique optical properties suitable for developing novel biosensing platforms.
Purpose of the Study:
- To develop a simple, sensitive, and label-free fluorescence strategy for the detection of trypsin (Try).
- To utilize the interaction between carbon quantum dots (CQDs) and protamine (Pro) for a "turn-on" fluorescence assay.
- To demonstrate the applicability of this method for real-world applications like trypsin monitoring and inhibitor screening.
Main Methods:
- Fabrication of carbon quantum dots (CQDs) and their complexation with protamine (Pro) leading to fluorescence quenching.
- Exploitation of trypsin's enzymatic activity to hydrolyze protamine, causing deaggregation of the CQDs/Pro complex.
- Restoration of CQDs fluorescence upon deaggregation, enabling a "turn-on" detection mechanism.
Main Results:
- A "turn-on" fluorescence assay for trypsin was successfully developed based on CQDs/Pro aggregation and deaggregation.
- The assay exhibited a linear response to trypsin concentration in the range of 25–500 ng mL⁻¹, with a limit of detection (LOD) of 8.08 ng mL⁻¹.
- The method demonstrated high specificity, convenience, and robustness against interfering substances, with successful application in real samples.
Conclusions:
- The developed fluorescence strategy provides a facile and effective method for trypsin detection.
- This CQDs-based assay holds significant potential for clinical applications, including disease diagnostics and high-throughput screening of trypsin inhibitors.
- The simplicity and sensitivity of the assay highlight the promise of nanomaterials in advanced analytical techniques.
