Figure 5.

Glycosylation remodeling of IgG using Endo-S and Endo-M-N175Q. (A) Schematic representation of the chemoenzymatic glycoengineering method. SG-Ox, sialoglycan-oxazoline. (B) SDS-PAGE of the transglycosylation product of IgG. Mogamulizumab (1 mg) was deglycosylated with Endo-S (2,000 U, New England Biolabs) in 50 mmol/L acetate containing 5 mmol/L CaCl₂ (pH 5.5) and purified on a protein G column. The deglycosylated IgG (0.5 mg) was incubated with Endo-M-N175Q (100 mU, Tokyo Chemical Industry, Japan) and 1 mg of SG-Ox (Fushimi Pharmaceutical, Japan) in 50 μL of 50 mmol/L sodium phosphate (pH 6.5) at 30°C for 2 h. CBB, Coomassie brilliant blue; SNA, Sambucus nigra agglutinin