A Novel Strategy for Characterization of Glycosylated Proteins Separated by Gel Electrophoresis.

Martin Larsen, Peter Skottrup, Jan Johannes Enghild, Peter Højrup, Peter Roepstorff

    Research output: Contribution to conferencePosterResearchpeer-review

    Abstract

    Protein glycosylation can be vital for changing the function or physiochemical properties of a protein. Abnormal glycosylation can lead to protein malfunction, resulting in severe diseases. Therefore, it is important to develop techniques for characterization of such modifications in proteins at a sensitivity level comparable with state-of-the-art proteomics. Whereas techniques exist for characterization of high abundant glycoproteins, no single method is presently capable of providing information on both site occupancy and glycan structure on a single band or spot excised from an electrophoretic gel. We present a new technique, which allows full characterization of low amounts of glycoproteins separated by gel electrophoresis. The method takes advantage of sequential specific and non-specific enzymatic treatment, followed by selective purification and characterization of the glycopeptides using graphite powder micro-columns in combination with mass spectrometry. The method is faster and more sensitive than previous approaches and would be ideal for proteomics studies and verification of correct glycosylation of recombinant glycoproteins.
    Original languageEnglish
    Publication date2005
    Publication statusPublished - 2005
    Event52nd ASMS Conference on Mass Spectrometry and Allied Topics - Nashville, TN, USA
    Duration: 1 Jan 2004 → …

    Conference

    Conference52nd ASMS Conference on Mass Spectrometry and Allied Topics
    CityNashville, TN, USA
    Period01/01/2004 → …

    Cite this

    Larsen, M., Skottrup, P., Enghild, J. J., Højrup, P., & Roepstorff, P. (2005). A Novel Strategy for Characterization of Glycosylated Proteins Separated by Gel Electrophoresis.. Poster session presented at 52nd ASMS Conference on Mass Spectrometry and Allied Topics, Nashville, TN, USA, .