Surface-enhanced Raman scattering-based detection of plasmin activity by specific peptide substrate

In this study, a new surface-enhanced Raman scattering (SERS)-based method has been developed for the detection of plasmin activity. Firstly, different peptide sequences, which are specific to plasmin, were examined. Then, SERS substrates were prepared by chosen peptide substrate. Enzyme activity was determined by pursuing the reduction of DTNB band at 1331 cm-1 with Raman spectroscopy. The reduction in SERS intensity was related to the plasmin activity, and changes in SERS intensity vs. plasmin concentration graph was obtained. Limit of detection (LOD) and limit of quantification (LOQ) values were calculated as 2.14 U/mL and 6.42 U/mL, respectively. Intra-and inter-day repeatability results were determined as 1.45% and 1.47% relative standard deviation (RSD). Also, recovery of the method was determined for the plasmin spiked milk samples. The results demonstrated that the proposed method could be successfully used to detect the plasmin activity in milk samples.

Automated summary

A new SERS-based method was developed for detecting plasmin activity, using specific peptide sequences and SERS substrates to determine enzyme activity and obtaining a graph of plasmin concentration versus SERS intensity changes. LOD and LOQ values were calculated, and the repeatability and recovery of the method were tested for plasmin spiked milk samples.