Fluorescence Assay for Phospholipase C Activity Using Liposome Probes / 分析化学

ABSTRACT

A simple assay for detection of phospholipase C (PLC) activity was developed based on a fluorescence liposome probe using the Liss Rhod PE-loaded phospholipid liposomes.The liposome probe was prepared by the coassembly of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and fluorescent lipid (Liss Rhod PE).The probe showed very low background fluorescence due to fluorescence self-quenching effect of Liss Rhod PE.As the PLC enzyme selectively digested lipid, the Rhod fluorescence was recovered from its quenched state, leading to the sensitive detection of PLC.This assay provided a limit of detection (at a signal-to-noise ratio of 3) of 2 U/L for PLC.In the presence of PLC inhibitor, the fluorescent response of the sensor for PLC decreased, indicating that the assay could also be used for screening PLC inhibitors.