O-glycosylation has important biological function and is therefore key in research of disease and development of biological treatment for it. There are two ways to determine binding site of O-glycans to proteins. The first is release of O-glycans with enzymatic and chemical methods, following treatment to more stable and labeled derivates before analysis. The second is decomposition of O-glycoprotein, where glycans stay intact and binded to O-glycopeptides. After that enrichment, separation and analysis with different types of fragmentation in MS are done. I developed a method for separation of fluorescent-labelled O-glycans with liquid chromatography in experimental part. I first developed method on HILIC column on maltodextrins, maltose and glucose, and then used it on standard solution because of high prices. Excess of label reagent resulted in bad separation, so I removed it with SPE extraction. Later I also used mixed mode chromatography. After few optimization steps this method offered better results. During validation I checked linearity, limit of detection, limit of quantification and repeatability.
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