Fluorescent microsphere probe for rapid qualitative and quantitative detection of trypsin activity

Abstract

Current technologies still face a big challenge to achieve simultaneous rapid qualitative and quantitative detection of trypsin. In our present study, we developed a simple and effective strategy to sensitively, qualitatively and quantitatively analyze the activity of trypsin using a fluorescent polystyrene (PS) microsphere probe. PS spheres were first functionalized by the surface coating of polyethylene glycol (PEG), which could significantly decrease the possibility of nonspecific physical adsorption of the fluorescein isothiocyanate isomer-modified peptide (peptide-FITC). Then, the obtained PS-PEG spheres were chemically interacted with peptide-FITC, which were then employed to monitor the real-time activity of trypsin. The peptide used in our work contained rich lysine and arginine residues, which were the recognition sites of trypsin. When trypsin interacted with the PS-FITC-peptide microspheres, the peptide-FITC rapidly decomposed into free small fragments in solution, resulting in a gradual decrease in the fluorescence of the PS spheres. By taking advantage of the fluorescence changes using confocal microscopy imaging and fluorescence spectrum intensity, it is easy to achieve the qualitative and quantitative detection of trypsin, with a highly sensitive detection limit as low as 0.5 ng mL⁻¹ and high selectivity. Thus, the designed fluorescent PS microsphere probe would be very promising in various applications such as food safety inspection, personal healthcare and on-site environmental monitoring.