Heparin-enhanced peroxidase-like activity of iron-cobalt oxide nanosheets for sensitive colorimetric detection of trypsin

Iron-cobalt oxide nanosheets (FeCo-ONSs) were proved to have intrinsic peroxidase-like activity. Additionally, the peroxidase-like activity of FeCo-ONSs toward the oxidation of 3,3,5,5-tetramethylbenzidine (TMB) was dramatically enhanced after heparin addition due to the stronger affinity toward TMB. Protamine combines with heparin, so the promotion of peroxidase-like activity of FeCo-ONSs with heparin was suppressed. With the addition of trypsin, protamine was hydrolyzed and the enhancement effect of catalytic activity of FeCo-ONSs was recovered. Based on above process, a sensitive colorimetric platform for trypsin activity determination was constructed through measuring the absorbance of produced oxTMB at 652 nm, providing a linear detection range of 5 to 500 ng/mL and a low detection limit of 2.8 ng/mL. The method was applied to trypsin determination in real samples (human urine sample and multienzyme tablet sample) with satisfactory results, illustrating the potential application of this biosensor.