GlycINATOR (EndoS2) is an IgG-specific endoglycosidase that hydrolyzes all glycoforms present at the Fc-glycosylation site. The enzyme acts on the chitobiose core and leaves the core GlcNAc.
GlycINATOR is available in three different product formats: as lyophilized powder (GlycINATOR Enzyme), as lyophilized powder with low levels of endotoxin (GlycINATOR LE), and immobilized on agarose (Immobilized GlycINATOR).
GlycINATOR (EndoS2) is an endoglycosidase from Streptococcus pyogenes that specifically hydrolyzes glycans at the Fc glycosylation site of IgG. The enzyme has been demonstrated to remove all glycoforms on IgG, including high-mannose, hybrid, complex, and bisecting type glycans (Sjögren et al. 2015). GlycINATOR deglycosylates all human IgG subclasses and IgG from many different species, including mouse, rat, monkey, sheep, goat, cow, and horse. It has been reported to have limited activity on glycans from alpha-1-acid glycoprotein (Sjögren et al. 2013). The enzyme is expressed in E. coli, contains a His-tag, and the molecular weight is 92 kDa.
The enzyme is used to reduce the complexity of antibody based therapeutics or to study core afucosylation and N-glycan site occupancy. The rapid and mild reaction conditions of GlycINATOR ensure a minimized risk for sample prep-induced modifications (Sjögren et al. 2015).
Sjögren, J. et al., 2013. EndoS2 is a unique and conserved enzyme of serotype M49 group A Streptococcus that hydrolyses N-linked glycans on IgG and α1-acid glycoprotein. The Biochemical Journal, 455(1), pp.107–118.
Sjögren, J. et al., 2015. EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. Glycobiology, 25(10), pp.1053–1063.
- Cleaves all N-linked glycans on IgG
- Specific for the Fc-glycosylation site
- Works on native IgG
- Rapid deglycosylation - 30 min protocol
- Level of afucosylation
Afucosylation of the IgG glycans has substantial impact on the effector functions of the antibody and can be studied using GlycINATOR.
- Reduce Sample Complexity
The glycans can quickly be removed from native IgG using GlycINATOR for subsequent intact mass analysis.
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GlycINATOR enzyme supplied as lyophilized powder. The enzyme is available in 2000 unit vials for deglycosylation of up to 2 mg IgG.
GlycINATOR enzyme lyophilized for endotoxin-sensitive assays. This product formulation has low levels of endotoxins per vial (< 0.2 EU). Available in 2000 unit vials.
GlycINATOR enzyme immobilized on agarose, allowing for IgG deglycosylation without contaminating the final preparation with GlycINATOR enzyme.
We have developed an “Enzyme Reference Tool” that organizes all published papers using our products. With this tool, you can filter by quality attribute, enzyme used, type of antibody molecule, and technology.
Latest reference using this product
- Orthogonal liquid chromatography–mass spectrometry methods for the comprehensive characterization of therapeutic glycoproteins, from released glycans to intact protein level
Eric Largy, Fabrice Cantais, Géry Van Vyncht, Alain Beck, Arnaud Delobel
- Orthogonal Assessment of Biotherapeutic Glycosylation: A Case Study Correlating N-Glycan Core Afucosylation of Herceptin with Mechanism of Action
Rosie Upton, Leonard Bell, Colin Guy, Paul Caldwell, Sian Estdale, Perdita E. Barran and David Firth
Patent and Disclaimer
The trademark GlycINATOR is the property of Genovis AB.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.