Ultrasensitive detection and quantification of acidic disaccharides using capillary electrophoresis and quantum dot-based fluorescence resonance energy transfer.

摘要: Rapid and highly sensitive detection of the carbohydrate components glycoconjugates is critical for advancing glycobiology. Fluorescence (or Forster) resonance energy transfer (FRET) commonly used in DNA, protein structural biology, protease assays but less frequently applied to glycan analysis due difficulties inserting two fluorescent tags into small structures. We report an ultrasensitive method quantification a chondroitin sulfate disaccharide based on FRET, involving CdSe-ZnS core-shell nanocrystal quantum dot (QD) streptavidin conjugate donor Cy5 acceptor. The was doubly labeled with biotin Cy5. QDs then served concentrate target disaccharide, enhancing overall efficiency, unlinked hydrazide producing nearly zero background signal capillary electrophoresis using laser-induced fluorescence different band-pass filters. This generally applicable acidic glycans offers promise high-throughput glycosaminoglycans.