Does GalactEXO digest fucosylated glycan structures such as Lewis X or Lewis a?

No. Branching fucose linked to the same residue as the β-galactose inhibits GalactEXO activity and needs to be removed prior to GalactEXO treatment. The FucosEXO enzyme can be used for hydrolysis of α1-2, α1-3 and α1-4 linked fucose.

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How long is the reaction time for GalactEXO?

For most substrates such as antibodies or Fc-fusion proteins a 2 h incubation results in hydrolysis of over 95% of the present galactose. More complex substrates with high degree of galactosylation may require longer incubation times.

Does GalactEXO hydrolyze galactose on N and O-glycosylated proteins?

Yes, both beta1-3 linked galactose on O-glycans and beta1-4 galactose on N-glycans are hydrolyzed by GalactEXO.

Can you combine GalactEXO with SialEXO or SialEXO 23 to remove both galactoses and sialic acids in a single reaction?

Yes, you can. All three enzymes perform well in the same reaction buffer (20 mM Tris pH 6.8). There is no need to increase incubation time when combining the enzymes.

Can I use SialEXO in combination with OglyZOR or OpeRATOR?

Yes, SialEXO can be incubated together with OpeRATOR and/or OglyZOR in 20 mM Tris pH 6.8.

Can I use SialEXO in combination with PNGaseF?

Yes, both enzymes are active under native conditions, preferably use 20 mM Tris pH 7. If denaturing reaction conditions are required for PNGaseF, the reaction with SialEXO needs to be performed first under native conditions, then add PNGaseF and the required buffer.

Does SialEXO contain a purification tag?

Yes, SialEXO contains a 6x His-tag.

Does SialEXO require denaturation of the proteins for activity on glycoproteins?

No, SialEXO works both on glycoproteins in native conditions but can also be used on denatured proteins after buffer exchange.

Can I use SialEXO for desialylation of both N- and O-glycans?

Yes, SialEXO displays activity on sialic acids with α2-3, α2-6 and α2-8 linked bonds of both N- and O-glycans.

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