| Home > Departments > Diagnostic Center > Department of Clinical Chemistry/Central Laboratories > Research topics, projects & Methods > Glycoproteomics by affinity chromatography and quantitative mass spectrometry
Protein glycosylation is a key determinant of cell recognition. Also, changes in protein glycosylation have been shown to be associated with a number of pathological states, including cancer (1,2).
In the ongoing project, we use the affinity of various lectins for specific glycoprotein structures in order to fractionate the highly complex proteome of the humane plasma. By isolating the the fraction specifically bound to a particular lectin we deplete the samples of all proteins lacking the sugar structure recognized by the respective lectin. Further depletion of abundant proteins by immunoaffinity chromatography enables the detection of proteins of very low abundance, which may function as diagnostic or prognostic biomarkers.
Furthermore, we are using the differential affinity of lectins as a tool for glycoprotein profiling in order to detect changes in glycoprotein structure associated with desease states. At present, we apply plant lectins for this purpose, but at a later stage we plan to use mammalian glycoreceptor proteins in these experiments, which would give a more realistic insight into the changes in cell-cell-interactions triggered by changes in the glycosylation pattern of cells surface proteins. Tandem mass spectrometry, in combination with enzymatic techniques, is also be used to elucidate the structure of the glycan moieties involved in these interactions. An example of the application of ESI-QTOF tandem mass spectrometry for the analysis of glycan structures is given in ref. (3).
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