GlycINATOR® (EndoS2)
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 intact.
GlycINATOR is available in three different product formats:
- GlycINATOR Enzyme - lyophilized enzyme
- GlycINATOR LE - lyophilized enzyme with low levels of endotoxin
- Immobilized GlycINATOR - immobilised on agarose beads
Product Information
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.
Key Characteristics
- Human IgG1-4, Fc-fusion proteins, IgG from mouse, rabbit, rat, monkey, sheep, goat, cow and horse
- 30 min reaction
- Requires native IgG fold
- All Fc glycoforms of IgG
Applications
- 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. - GlyCLICK
Site-specific conjugation of native IgG using enzymatic remodelling of the Fc glycans
Find all applications here
Product Formats
GlycINATOR® Enzyme
GlycINATOR enzyme supplied as lyophilized powder. The enzyme is available in 2000 unit vials for deglycosylation of up to 2 mg IgG.
GlycINATOR® LE
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.
Immobilized GlycINATOR®
GlycINATOR enzyme immobilized on agarose, allowing for IgG deglycosylation without contaminating the final preparation with GlycINATOR enzyme.
Latest References using this Product
- Protocols for the analytical characterization of therapeutic monoclonal antibodies. II – Enzymatic and chemical sample preparation
2017-06-19
Balazs Bobaly, Valentina D’Atri, Alexandre Goyon, Olivier Colas, Alain Beck, Szabolcs Fekete, Davy Guillarme - Monitoring Glycosylation Profile and Protein Titer in Cell Culture Samples Using ZipChip CE-MS
2017-04-08
Yan Wang, Peng Feng, Zoran Sosic and Li Zang
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Product Documents
Patent and Disclaimer
GlycINATOR®
All rights reserved. Aspects of GlycINATOR technology are encompassed by pending patent applications and by granted Patent in USA, EPO, Singapore and China in the name of Genovis AB. The trademark GlycINATOR is the property of Genovis AB.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.