prikaz prve stranice dokumenta Mouse Fc and Fab IgG N-glycosylation
  Download
PDF 1.56 MB
master's thesis
Mouse Fc and Fab IgG N-glycosylation
Zagreb: University of Zagreb, Faculty of Science, 2021. urn:nbn:hr:217:427228
Vuković, Nikolina
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Vuković, N. (2021). Mouse Fc and Fab IgG N-glycosylation (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:427228

Vuković, Nikolina. "Mouse Fc and Fab IgG N-glycosylation." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:427228

Vuković, Nikolina. "Mouse Fc and Fab IgG N-glycosylation." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:427228

Vuković, N. (2021). 'Mouse Fc and Fab IgG N-glycosylation', Master's thesis, University of Zagreb, Faculty of Science, accessed 27 December 2024, https://urn.nsk.hr/urn:nbn:hr:217:427228

Vuković N. Mouse Fc and Fab IgG N-glycosylation [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2021 [cited 2024 December 27] Available at: https://urn.nsk.hr/urn:nbn:hr:217:427228

N. Vuković, "Mouse Fc and Fab IgG N-glycosylation", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:217:427228

Metadata
TitleMouse Fc and Fab IgG N-glycosylation
Title (english)Mouse Fc and Fab IgG N-glycosylation
AuthorNikolina Vuković
MentorMarija Pezer (mentor)
MentorSofia Ana Blažević (sumentor)
Committee memberSofia Ana Blažević (predsjednik povjerenstva)
Committee memberJasna Lajtner (član povjerenstva)
Committee memberPetra Peharec Štefanić (član povjerenstva)
Committee memberAna Galov (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2021-02-24, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
N-glikani su kompleksni oligosaharidi vezani na dušikov atom asparagine. Imunoglobulin G (IgG) sadrži dva N-glikana vezana na kristalizirajućem fragmentu (Fc). Manji udio molekula IgG ima vezane N-glikane na antitijelo vezujućem fragment (Fab). Fab i Fc N-glikani utječu na funkciju molekule IgG u imunološkom odgovoru, što se često istražuje na mišjim modelima. Cilj ovog diplomskog rada je bio uspostaviti analitičku metodu za usporednu analizu N-glikozilacije Fc i Fab fragmenta mišjeg imunoglobulina G na temelju metode koja se koristi za analizu glikoma ljudskog IgG-a. IgG je izoliran afinitetnom kromatografijom iz seruma miševa soja CBAT6T6. Optimirani protokol uključuje dvadesetsatnu razgradnju na kuglicama specifičnim za Fc fragment u reducirajućim uvjetima. Nakon razgradnje na Fab i Fc fragment, uzorci su deglikozilirani. Oslobođeni N-glikani su fluorescentno obilježeni i analizirani kapilarnom gel-elektroforezom s laserom induciranom fluorescencijom u DNA sekvenceru. Prikupljeni podaci su obrađeni u programu Empower. Ukupno je određeno 35 N-glikanskih pikova, od kojih je za njih 16 otprije poznat N-glikanski sastav. Inicijalno upotrijebljena metoda za analizu N-glikana Fab i Fc fragmenta mišjeg IgG-a procijenjena je učinkovitom. Usporedba postotaka ukupne visine (PHP%) pojedinih pikova pokazala je razlike u glikanskom profilu Fc i Fab fragmenata nalik onima uočenim kod ljudskog IgG-a.
Abstract (english)
N-glycans are complex oligosaccharides bound to the nitrogen atom of asparagine. Immunoglobulin G (IgG) molecule contains two N-glycans attached to the fragment crystallizable (Fc) region, but only a minority of IgG molecules contain antigen binding fragment (Fab) N-glycans. Both Fab and Fc N-glycans influence the molecule's function in the immune response, a phenomenon often explored through mouse models. The aim of this study was to establish an analytical method for comparative N-glycosylation analysis of mouse Fc and Fab IgG fragments based on the method used for human IgG. IgG was isolated by affinity chromatography from serum of CBAT6T6 mice. The optimized protocol included a 20h-digestion on Fc beads in reducing conditions and fragments separation. Each IgG portion was deglycosylated and the released N-glycans were fluorescently labelled and analysed by capillary gel-electrophoresis with laser-induced fluorescence on an DNA sequencer. Collected data were processed in the Empower program. A total of 35 N-glycan peaks were defined and 16 of them annotated. The initially implemented method for the analysis of mouse Fab and Fc IgG N-glycans is judged functional. The comparison of proportion height percentages (PHP%) showed differences in the Fab and Fc glycoprofile similar to those observed for human IgG.
Keywords
Keywords (english)
Languageenglish
URN:NBNurn:nbn:hr:217:427228
Study programmeTitle: Experimental Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra eksperimentalne biologije (magistar/magistra eksperimentalne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2023-02-24
Terms of useLicence In copyright icon
Created on2021-03-08 14:12:12