Combining 3D graphene-like screen-printed carbon electrode with methylene blue-loaded liposomal nanoprobes for phospholipase A2 detection

Phospholipase A2 (PLA2) enzyme could be acted as a unique biomarker for forecasting and diagnosing certain diseases. Therefore, it is important to monitor PLA2 activity in biological and clinical samples. In this work, a simple electrochemical assay for PLA2 activity was developed based on a screen-printed carbon electrode (SPCE) with 3D graphene-like surface. When the PLA2-containing sample was mixed with the nanoprobes, i.e. the electroactive marker methylene blue (MB) encapsulated within nanometer-sized phospholipid liposomes, MB was released and adsorbed/enriched in site onto the surface of SPCE in a micro-cell. The encapsulation and enzymatic release of MB were evaluated using UV-Vis and fluorescence. The peak current due to oxidation of the adsorbed MB on the SPCE was measured by square-wave voltammetry (SWV). The current was directly linear to the PLA2 activity from 5 U/L to 200 U/L with a detection limit of 3 U/L. The same method can also be used for screening PLA2 inhibitors. Thus, the enrichment strategy developed in this work could be a promising signal amplification method for the sensitive and selective detection of PLA2 in biological or clinical samples.