Fast and sensitive detection of protein concentration in mild environments

Determination of protein concentration in mild environments is of great significance in the clinic diagnose and bioassay. Herein, a simple, fast and sensitive method for protein quantitative determination in neutral solution (pH 7.0) is developed. This assay is based on competition adsorption of the sample protein and fluorescently labeled dog serum albumin (FITC-DSA) onto gold nanoparticles (AuNPs). As the competitor FITC-DSA molecules are added into the mixture solution of sample protein conjugated AuNPs, they will compete for active sites of AuNPs, resulting in decrease in fluorescence intensity due to the quenching effect of AuNPs via fluorescence resonance energy transfer (FRET). Thus, quantitative determination of sample protein concentration can be achieved. Under the optimum conditions, the decrease in fluorescence intensity of the solution is related to the concentration of sample protein and a low detection limit of 0.01 µg/mL BSA can be achieved in 5 min. For the validation of our strategy in practical applications, the total protein content in human serum was determined using the as-proposed method. The result is in well agreement with that of measured by other conventional methods, suggesting a simple, accurate, and mild approach for protein detection in bioassay.