Fast and Easy Detection System of Protein Glycosylation by Using Fluorescence Resonant Energy Transfer between Nanoparticles

摘要

We construct the new glycosylation detection system using Fluorescence Resonance Energy Transfer (FRET) between gold nanoparticles (AuNPs) and semiconductor quantum dots (QDs). FRET can be induced by biomolecular interactions between concanavalin A (conA)-conjugated AuNP (conA-AuNP) as an acceptor and dextran-conjugated QD (dex-QD) as a donor, resulting the photoluminescence (PL) quenching of QDs. The FRET efficiency was changed in the presence of glycoprotein that competes with the dextran on dex-QDs on the binding site of conA-AuNPs. The inhibition effect is concentration dependent. And using this NPs-based FRET system we can detect the deglycosylation and neoglycosylation states of protein using Avidin, NeutrAvidinTM, Bovine Serum Albumin (BSA) and diversely neoglycosylated BSA. Additionally we applied this system to discriminate the real glycoprotein products, recombinant glucose oxidase (rGOD) from yeas and fungus with varied branch types of mannosylation. We reported FRET based inhibition assay using QDs and AuNPs applied to the model system, streptavidin-biotin system.1 Now further studying results will be suggested about glycosylation detection with more efficient and real product system, using previous accomplishment. This easy, fast and non labeling method showed the possibility to open new high-throughput detection method for the screening glycoprotein-based drugs.