Razvoj metode za ukoncentriravanje N-glikana PGC ekstrakcijom na čvrstoj fazi

master's thesis

• Title (english): Development of the method for N-glycan enrichment using PGC solid phase extraction
• Author: Isabella Koprivec
• Mentor: Toma Keser (mentor)
• Committee member: Toma Keser (predsjednik povjerenstva)
• Committee member: Kristina Pavić (član povjerenstva)
• Committee member: Anita Hafner (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of biochemistry and molecular biology); Zagreb
• Defense date and country: 2018-09-27, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

N-glikom predstavlja jedinstvenu poveznicu između genskih predispozicija za razvoj
pojedinih bolesti i okolišnih faktora kojima je organizam izložen te je stoga moguća njegova uporaba
kao biomarkera u preventivnoj i personaliziranoj medicini. Jedan je od glavnih ciljeva moderne
glikobiologije razviti brze i osjetljive metode za analizu N-glikana u tragovima. Razvijenom metodom
za ukoncentriravanje N-glikana PGC ekstrakcijom na čvrstoj fazi moguće je pomoću HILIC-UPLCFLR kvantitativno odrediti 5x manje koncentracije N-glikana u odnosu na prijašnju metodu. Kako bi
kromatografski profil bio prihvatljiv, tijekom analize potrebno je pripaziti na kompatibilnost reagensa
s mobilnom fazom za kromatografsku analizu, ionsku jakost prilikom eluacije glikana s PGC-a i udio
vode u eluatu glikana. Razvijena metoda ima potencijal korištenja u istraživanjima različitih stanja i
bolesti kod kojih dolazi do promjene glikozilacijskog profila, a u kojima je potrebna veća osjetljivost
zbog male količine glikana u uzorku.

Abstract (english)

N-glycome represents a unique connection between the genetic predisposition for the
development of certain diseases and the environmental exposure of the organism, making it a
potential biomarker in preventive and personalised medicine. Consequently, the development of fast
and sensitive methods for the analysis of trace N-glycans is one of the main goals in the field. By
using the newly developed PGC SPE method for the enrichment of N-glycans, followed by HILICUPLC-FLR, it is possible to quantitatively measure 5x lower concentrations of N-glycans than with
previous methods. During the analysis, it is important to adjust the compatibility of the reagents with
the mobile phase used in chromatography, ionic strength for eluation of the N-glycans from PGC and
the amount of water in the N-glycan eluate. The method can be used in research of different
conditions and diseases which are known to have an altered glycosylation profile, as well as in
molecular diagnostics.

• Keywords: N-glikani; koncentriranje; PGC; HILIC-UPLC
• Keywords (english): N-glycans; enrichment; PGC; HILIC-UPLC
• Language: croatian
• URN:NBN: urn:nbn:hr:163:654660
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2020-02-18 16:24:38