FAQ: Which enzyme should be used to remove β1-4 linked Galactose residues from intact IgG?

β1-4 linked galactose residues can be difficult to remove from intact IgG under native conditions, and the activity varies greatly depending on the subtype of IgG. Extended incubations and increased units are usually needed to see substrate turnover. β1-4 Galactosidase S (NEB# P0745) is the preferred enzyme for this purpose as β1-4 Galactosidase (NEB# P0730) has very low activity on intact IgG. A protocol using up to 50*g of Rituximab (CHO derived IgG1) with 80 units (10μl) of β1-4 Galactosidase S (NEB# P0745) in 1X GlycoBuffer 1 in a total volume of 50μl, incubated for 24 hours at 37°C yields efficient removal of β1-4 galactose residues. However, similar conditions treating a murine IgG2a yields only ~70-80% removal of β1-4 galactose residues.