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Dissertationen online der Freien Universität Berlin
|Haupttitel||Ion Mobility-Mass Spectrometry of Complex Carbohydrates|
|Titelvariante||Ionenmobilitäts-Massenspektrometrie von komplexen Kohlenhydraten|
Geburtsort: Berlin, Deutschland
|Gutachter||Prof. Dr. Kevin Pagel|
|weitere Gutachter||Prof. Dr. Christoph Schalley|
|Freie Schlagwörter||ion mobility; mass spectrometry; glycans; carbohydrates|
|DDC||500 Naturwissenschaften und Mathematik|
543 Analytische Chemie
|Zusammenfassung||Carbohydrates, also referred to as glycans or oligosaccharides, are biomolecules that play an important role in all living organisms. They are, for example, involved in cell-cell recognition, protein folding or the development of cancer. Oligosaccharides consist of monosaccharide building blocks, which are often isomers that only differ in the stereochemistry at individual carbons. Additionally, they contain several hydroxyl groups that can be connected to other monosaccharides via the formation of a glycosidic bond. This often results in branched structures with a complex regio- and stereochemistry. The high structural diversity of glycans provides an enormous challenge for almost all areas of the glycosciences.|
A promising technique for the structural investigation of carbohydrates is ion mobility-mass spectrometry (IM-MS). IM-MS measures the mass-to-charge ratio (m/z) of ions as well as the time they need to traverse a cell, filled with inert gas, under the influence of a weak electric field. Ions are separated according to their mass, charge, size, and shape, which enables the differentiation of isomers. In addition, an obtained drift time can be converted into a collision cross section (CCS), which is a molecular property that can be universally compared.
In this thesis, the potential of IM-MS for the structural analysis of glycans was investigated. First, a variety of N-glycans and the polysaccharide dextran were examined and the absolute CCSs of their intact ions and fragments reported. Subsequently, these reference values were used to establish a CCS calibration procedure for commercial traveling wave IM-MS instruments. In addition, the influence of the utilized drift gas and the ion’s charge on the quality of a calibration were investigated. To assess the capability of IM-MS, a systematic study was performed using a set of six synthetic carbohydrates that represent all possible types of isomerism. It was shown that regio- and stereoisomers can be identified and separated when analyzed as deprotonated ions. Furthermore, the relative concentration of an isomer in a mixture was determined. With the increasing size of glycans it becomes exceedingly more difficult to identify small structural differences. In this thesis, it was shown that fragments, which contain two to five monosaccharides, are often more informative than their precursor ions. This can be exploited to identify common glycan motifs within larger glycans, based on characteristic fragments with unique CCSs. It was, for example, shown that blood group epitopes can function as such marker fragments. In addition, N-acetylneuraminic acids, linked either via an α2→3 or α2→6 glycosidic bond to larger glycans, were identified using characteristic fragments. The application of this approach is independent of the type of precursor and was successfully applied to N-glycans, milk sugars and glycopeptides.
Overall, IM-MS can be used to generate dual sets of m/z and CCS information, which form unique signatures that enable a fast and accurate identification of glycans. The data are ideally suited to be implemented into databases and provide the basis for the future development of automated, high-throughput glycan analysis.
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|Seitenzahl||XIV, 119 Seiten|
|Fachbereich/Einrichtung||FB Biologie, Chemie, Pharmazie|
|Tag der Disputation||26.09.2017|
|Erstellt am||10.10.2017 - 06:21:51|
|Letzte Änderung||11.10.2017 - 12:09:54|