IgGZERO® (EndoS)
IgGZERO (EndoS) is an IgG-specific endoglycosidase acting on complex type N-glycans at the Fc-glycosylation site of IgG. The enzyme acts on the chitobiose core and leaves the core GlcNAc intact.
Available in three different product formats:
- IgGZERO Enzyme - lyophilized enzyme
- IgGZERO LE - lyophilized enzyme with low levels of endotoxin
- deGlycIT - immobilized on agarose beads
Product Information
IgGZERO (EndoS) is an endoglycosidase from Streptococcus pyogenes that specifically hydrolyzes N-linked glycans in the Fc-region of native IgG. It can be used to rapidly deglycosylate antibodies to reduce sample complexity, or to inactivate Fc-mediated effector functions.
The Enzyme deglycosylates all human IgG subclasses, and IgGs from many different species, including mouse, rat, monkey, sheep, goat, cow, and horse. The enzyme only hydrolyzes glycans from the Fc-domain of IgG, even when other glycosylated proteins are present in the solution. The enzyme is expressed in E. coli, contains a His-tag, and the molecular weight is 110 kDa.
The IgGZERO enzyme has limited activity on high-mannose and hybrid-type glycans (Sjögren et al., 2015). If you are interested in endoglycosidases that hydrolyze all glycoforms on IgG, we recommend the GlycINATOR product family.
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, rat, monkey, sheep, goat, cow and horse
- 30 min reaction
- Requires native IgG fold
- Complex Fc glycoforms, limited activity on high-mannose and hybrid-type glycans
Applications
- Level of afucosylation
Afucosylation of the IgG glycans has substantial impact on the effector functions of the antibody and IgGZERO can be applied to study the degree of core fucosylation. - Inactivating antibodies
Removal of IgG glycans using IgGZERO has been proposed to reduce antibody binding to its Fc receptors.
Find all applications here
Product Formats
IgGZERO® Enzyme
Enzyme supplied as lyophilized powder. The enzyme is available in 1000 and 5000 unit formats (1 unit digest 1 µg IgG).
IgGZERO® LE
Enzyme lyophilized for endotoxin-sensitive assays. This product formulation has low levels of endotoxins per vial (< 0.2 EU). Available in 2000 unit vials.
deGlycIT™
deGlycIT is IgGZERO enzyme immobilized on agarose allowing for antibody Fc deglycosylation without contaminating the final preparation with IgGZERO enzyme.
Product Documents
Brochures, Posters and Applications
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External References and Reference Tool
FAQ and Support
Popular FAQ
Yes, IgGZERO/deGlycIT are compatible with most commonly used buffers with pH ranging from 6.0 to 8.0, but the reaction conditions might need to be optimized to ensure optimal deglycosylation.
Yes, many buffers are compatible with IgGZERO/DeGlycIT. Please keep pH close to neutral.
First you could increase incubation time and increase temperature to 37°C. One explanation for incomplete deglycosylation with deGlycIT could be that the antibody has higher amounts of high mannose, hybrid glycans or bisected glycans. For these glycans IgGZERO has low or no activity. High mannose glycans are particularly resistant to digestion recommend you try our other endoglycosidase GlycINATOR (EndoS2). This enzyme has much higher activity on these types of glycans sometimes seen in mAbs.
Typically, GlycINATOR is the first choice since it displays enzymatic activity on all glycoforms present on IgG, including complex type, high mannose, bisected and hybrid type glycans. IgGZERO does not hydrolyze high mannose glycans. IgGZERO deglycosylates some species faster as compared to GlycINATOR, for example goat IgG.
IgGZERO (EndoS) and GlycINATOR (EndoS2) are both IgG-specific endoglycosidases that hydrolyses Fc glycans to the innermost GlcNAc. The GlycINATOR enzyme has a broader substrate selectivity and therefore much higher enzymatic activity on high mannose and some bisected and hybrid glycans that can occur on mAbs. IgGZERO deglycosylates some species faster as compared to GlycINATOR, for example goat IgG.
