Applying bimetallic gold/silver nanoclusters and gold nanoparticles for dairy trypsin and inhibitor analysis via inner-filter approach

Trypsin, a globular protein hydrolase, catalyzes the hydrolysis of peptide bonds at carboxyl groups of arginine and lysine residues, its detection holds paramount significance in the dairy industry, aiding in the diagnosis, management of animal diseases, and preservation of dairy products. This study introduces a highly sensitive fluorometric detection platform utilizing bimetallic gold/silver nanoclusters (Au/Ag NCs) and gold nanoparticles (AuNPs) in an "off-on-off" configuration for precise quantification of dairy trypsin and trypsin inhibitors. This fluorescence variation is leveraged to determine the concentration of trypsin and its inhibitors. Under optimal experimental conditions, the detection of trypsin demonstrates a linear range of 5 ∼ 2000 ng·mL −1 , a lowest detection limit of 1.18 ng·mL −1 , and an observed IC 50 of 5.94 μg·mL −1 for the soybean trypsin inhibitor (STI). Therefore, it could be inferred that the development of a fluorescence sensing platform presents an important basis for the detection of dairy trypsin.