Carbohydrate analysis for biomedical applications-developing integrated workflows for analysing glycans in neurodegenerative disorders

Sirathanarun, P. (2024) Carbohydrate analysis for biomedical applications-developing integrated workflows for analysing glycans in neurodegenerative disorders. PhD thesis, University of Reading. doi: 10.48683/1926.00128597

Abstract/Summary

This thesis aims to develop an analyHcal workflow uHlizing HPLC-fluorescence detecHon for studying N- and O-linked glycan levels in neurodegeneraHve disorders in vitro. This method was iniHally validated with simple model samples before being applied to complex N-glycan samples from a neuroblastoma cell line in vitro (Chapter 5). The in vitro applicaHon builds on method development for the various stages of the workflow, specifically the labelling (Chapter 2), release (Chapter 3) and enrichment (Chapter 4) of glycans and makes extensive use of HPLC. The synthesis of an alkyne reagent is described in Chapter 2, which also summarises its use in the CuAAC reacHon for successfully labelling O-glycans and in a reducHve aminaHon reacHon for successfully labelling N-glycans. The development of methods for simultaneously releasing O- and N-linked glycans from glycoproteins to allow their simultaneous analysis is described in Chapter 3. However, this could not be achieved despite extensive a;empts to achieve complete release of O- and N-linked glycans to monomer units. The enrichment of labelled N-glycans was also probed in Chapter 4, illustraHng that Amide SPE and co;on wool SPE demonstrated superior performance compared to other absorbents. The qualitaHve and quanHtaHve analysis of N-glycans in the SH-SY5Y cell line during differenHaHon (Chapter 5) using the developed workflow revealed altered expression of N-glycans during the differenHaHon process. These altered glycans can potenHally be explored as neurodegeneraHve disease biomarkers

Item Type	Thesis (PhD)
URI	https://centaur.reading.ac.uk/id/eprint/128597
Identification Number/DOI	10.48683/1926.00128597
Divisions	Life Sciences > School of Chemistry, Food and Pharmacy > School of Pharmacy
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