• Login
    View Item 
    •   DSpace@RPI Home
    • Rensselaer Libraries
    • RPI Theses Online (Complete)
    • View Item
    •   DSpace@RPI Home
    • Rensselaer Libraries
    • RPI Theses Online (Complete)
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

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

    Author
    Chang, Yuqing
    View/Open
    167146_Chang_rpi_0185E_10104.pdf (6.744Mb)
    Other Contributors
    Linhardt, Robert J.; Colón, Wilfredo; McGown, Linda Baine; Dordick, Jonathan S.;
    Date Issued
    2013-05
    Subject
    Chemistry
    Degree
    PhD;
    Terms of Use
    This electronic version is a licensed copy owned by Rensselaer Polytechnic Institute, Troy, NY. Copyright of original work retained by author.;
    Metadata
    Show full item record
    URI
    https://hdl.handle.net/20.500.13015/880
    Abstract
    Rapid and high-sensitive determination of carbohydrate components of glycoconjugates is critical for the advancement of glycobiology. However, conventional methods pose great challenges because of the low concentration of carbohydrate contents from many biological samples and lack of natural chromophores or fluorophores for robust analysis. Here we present two examples of using capillary electrophoresis (CE) coupled with laser-induced fluorescence (LIF) detection for ultrasensitive analysis of glycosaminoglycans (GAGs).; 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.; Quantitative disaccharide compositional analysis is one of the most widely used strategies for structural characterization of GAGs, which is directly related to understanding their biological functions. Current methods of HP/HS-derived, CS/DS-derived and HA-derived disaccharides analysis are completed in separate chromatographic or electrophoretic experiments, which calls for time-consuming multiple sample recoveries and separate analysis steps. To solve this problem, we developed a CE-LIF method to resolve seventeen AMAC-labeled GAG-derived disaccharides in a single run with attomolar detection level and applied it to biological samples. This method can also simplify conventional enzymatic degradation procedure.;
    Description
    May 2013; School of Science
    Department
    Dept. of Chemistry and Chemical Biology;
    Publisher
    Rensselaer Polytechnic Institute, Troy, NY
    Relationships
    Rensselaer Theses and Dissertations Online Collection;
    Access
    Restricted to current Rensselaer faculty, staff and students. Access inquiries may be directed to the Rensselaer Libraries.;
    Collections
    • RPI Theses Online (Complete)

    Browse

    All of DSpace@RPICommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsThis CollectionBy Issue DateAuthorsTitlesSubjects

    My Account

    Login

    DSpace software copyright © 2002-2022  DuraSpace
    Contact Us | Send Feedback
    DSpace Express is a service operated by 
    Atmire NV