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. 2013 Jun 7;135(26):9723–9732. doi: 10.1021/ja4014375

Figure 4.

Figure 4

Generation of differentially glycosylated IgG1 Fc. Normal-phase HPLC analysis of 2-AA-labeled N-linked glycans, released from target antibody glycoforms by in-gel PNGase F digestion. (A) Glycan profile of monoclonal IgG1 b12. (B) Glycan profile of IgG1 incubated with 50 U/mL Clostridium perfringens neuraminidase for 48 h at 37 °C. (C) Glycan profile of IgG1 incubated with 25 μg/mL β1,4-galactosyltransferase (B4GALTI) and 80 μM uridine 5′-diphosphogalactose in 50 mM HEPES, 10 mM MnCl2, pH 7.5 for 48 h at 37 °C. (D) Glycan profile of IgG1 sequentially treated with B4GALTI and α2,6-sialyltransferase I (ST6GALI) as described above.