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A Fluorescence-based Assay of Phospholipid Scramblase Activity
Published on: September 20, 2016
Laser desorption/ionization mass spectrometric assay for phospholipase activity based on graphene oxide/carbon
Jieon Lee1, Young-Kwan Kim, Dal-Hee Min
1Department of Chemistry, Korea Advanced Institute of Science and Technology, 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Korea.
Abstract:
A new method for quantitative phospholipase activity assays using mass spectrometry (MS) and a supported thin film consisting of a graphene oxide (GO)/carbon nanotube (CNT) double layer as a substrate for laser desorption ionization (LDI) has been developed. Phospholipids were very efficiently analyzed by LDI-time-of-flight (TOF) MS on the GO/CNT films, presumably because of the affinity of phospholipids for the GO/CNT surface. Therefore, the rate of lipid hydrolysis was conveniently measured using LDI-TOF mass spectra obtained from a GO/CNT surface on which the phospholipid hydrolysis reaction mixtures had been spotted by comparing the mass-peak intensities of reactants and products. The present platform for phospholipase assays based on MS and GO/CNT double-layer films enables quantitative measurements at low cost, allows assays to be performed in a short time, and is compatible with an array format, unlike conventional assay methods.
