Fast and Easy Detection System of Protein Glycosylation by Using Resonant Energy Transfer between Nanoparticles
E. Oh, D. Lee and H-S Kim
Korea Advanced Institute of Science and Technology, KR
FRET, gold nanoparticle, quantum dot, inhibition assay, glycoprotein
We construct the new glycosylation detection system using Resonance Energy Transfer (RET) between gold nanoparticles (AuNPs) and semiconductor quantum dots (QDs). RET 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 RET 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 RET 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.
This easy, fast and non labeling method showed the possibility to open new high-throughput detection method for the screening glycoprotein-based drugs.
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Nanotech 2006 Conference Program Abstract