Naslov
Mouse Fc and Fab IgG N-glycosylation

Autori
Vuković, Nikolina

Vrsta, podvrsta i kategorija rada
Ocjenski radovi, diplomski rad, diplomski

Fakultet
Prirodoslovno-matematički fakultet, Biološki odsjek

Mjesto
Zagreb

Datum
24.02

Godina
2021

Stranica
69

Mentor
Pezer, Marija ; Blažević, Sofia Ana

Ključne riječi
ragment crystallizable, antigen binding fragment, N-glycans, SDS-PAGE, CGE-LIF

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Temeljne medicinske znanosti

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