Fig. 3

IgG1-Fc produced as an N-GlcNAc modified glycoform. a Human IgG1-Fc expressed in P. pastoris strains and detected with Coomassie staining SDS-PAGE. Lanes 1–2: P. pastoris WT cultured for 3 days and 4 days; Lanes 3–4: P. pastoris MNS1-EndoT cultured for 3 days and 4 days; Lanes 5–6: P. pastoris MNN9-EndoT cultured for 3 days and 4 days; M stands for the protein marker. b IgG1-Fc purified from P. pastoris WT (Lane 1) and P. pastoris MNN9-EndoT (Lane 2). M stands for the protein marker. c MALDI-TOF MS analysis of peptide maps from digested recombinant IgG1-Fc proteins. The IgG1-Fc proteins secreted from E. coli (upper) and P. pastoris MNN9-EndoT (lower) were digested with Glu-C, and analyzed by MALDI-TOF MS. The peak with m/z at 2850.63 was assigned as the peptide (P295-QYNSTYRVVSVLTVLHQDWLNGKE-318), while the peak with m/z at 3053.66 was assigned as the peptide (P295–318) with a HexNAc moiety addition