Ultrasensitive analysis of glycosaminoglycans by capillary electrophoresis coupled with laser induced fluorescence detection

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Authors
Chang, Yuqing
Issue Date
2013-05
Type
Electronic thesis
Thesis
Language
ENG
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Chemistry
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Abstract
Fluorescence resonance energy transfer (FRET) is commonly used in DNA assays, in protease assays and in protein structural biology, but FRET is infrequently applied to carbohydrate analysis due to the lack of active sites on a carbohydrate molecule and proper pathways to assemble a FRET pair into a close proximity to produce efficient FRET. We developed a method based on a quantum dot (QD)-Cy5 hybrid FRET pair for carbohydrate analysis that is capable of detecting very low concentrations of disaccharides. QDs also function as a concentrator of target disaccharides and increase the overall FRET efficiency. Unlinked QDs and Cy5-hydrazide produce a near-zero background signal on CE-LIF with two different band-pass filters. With this simple model, we studied the quantitative properties of this FRET system. This method can be applied to ultrasensitive analysis of disaccharide analysis and potentially applied to that high-throughput analysis of GAGs.
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May 2013
School of Science
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Rensselaer Polytechnic Institute, Troy, NY
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