Strukturna analiza N-glikanskog profila staničnih linija HepG2 i 1.1B4 HILIC-UPLC-FLR-MS/MS metodom

master's thesis

• Title (english): Structural analysis of N-glycane profile in HepG2 and 1.1B4 cell lines using HILIC-UPLC-FLR-MS/MS method
• Author: Dinko Šoić
• Mentor: Toma Keser (mentor)
• Committee member: Toma Keser (predsjednik povjerenstva)
• Committee member: Biljana Nigović (član povjerenstva)
• Committee member: Tamara Pavić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of biochemistry and molecular biology); Zagreb
• Defense date and country: 2019-05-29, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Glikozilacija je najčešća i najkompleksnija modifikacija proteina te kao takva ima presudnu ulogu
na konačna svojstva i funkcije glikoproteina. Budući da se sinteza glikana ne odvija po predlošku
već je izrazito osjetljiva na okolišne uvjete i patofiziološke procese, dijagnostički potencijal
glikana sve se više istražuje. Zbog velike potrebe za jednostavnošću i lakoćom manipuliranja, kao
modeli u biomedicinskim istraživanjima naširoko se koriste stanične linije. Detaljna
karakterizacija njihovog glikanskog profila izrazito je važna za utvrđivanje uspješnosti korištenog
modela u glikomskim istraživanjima.
U ovom su radu analizirani N-glikani staničnih linija HepG2 i 1.1B4 HILIC-UPLC-FLR-MS/MS
metodom. Rezultati rada prikazani su asigniranim FLR kromatogramima i tablicama sa
strukturnim prikazom glikanskih pikova. U staničnoj liniji HepG2 utvrđena su 47 glikanska pika,
odnosno njih 46 u liniji 1.1B4. Glikanski profili staničnih linija uvelike su se preklapali, uz
nekoliko karakterističnih struktura za svaku liniju. Iako je većina struktura pripadala kompleksnom
tipu, najintenzivniji pikovi sadržavali su oligomanozni tip glikana. Dobiveni glikanski profili
razlikuju se od onih plazmatskih upravo po velikom udjelu oligomanoznog tipa N-vezanih glikana,
kao i po prisutnosti paucimanoznih struktura.

Abstract (english)

Glycosylation is the most common and complex modification of proteins and as such has a key
role in shaping the final properties and functions of glycoproteins. Since glycan synthesis is not
template-driven, but is instead strongly affected by environmental factors and pathophysiological
processes, the diagnostic potential of glycans is being thoroughly researched. Due to the strong
need for simplicity and ease of manipulation, cell lines are widely used as models in biomedicinal
research. Detailed characterization of their glycane profile is extremely important for determining
the success of the model used in glycome research.
This thesis analyzed N-glycans of HepG2 and 1.1B4 cell lines using HILIC-UPLC-FLR-MS/MS
method. The results are presented as assigned FLR chromatograms and tables with the structural
representation of the glycan peaks. In the HepG2 cell line there were 47 glycane peaks determined,
while in the 1.1B4 line there were 46 peaks respectively. The glycan profiles of the cell lines
vastly overlapped, with several characteristic structures for each line. Although most structures
belonged to a complex type, the most intense peaks contained a high-mannose type of glycans.
The obtained glycane profiles differ from those of the plasma precisely due to the large proportion
of high-mannose type of N-linked glycans, as well as the presence of the paucimannose
structures.

• Keywords: N-glikani; glikozilacija; stanične linije; HILIC-UPLC; MS/MS
• Keywords (english): N-glycans; glycosylation; cell lines; HILIC-UPLC; MS/MS
• Language: croatian
• URN:NBN: urn:nbn:hr:163:744165
• 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: 2020-02-24 10:37:31