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.
The IgGZERO enzyme is available in three different product formats: as lyophilized powder (IgGZERO Enzyme), as lyophilized powder with low levels of endotoxins (IgGZERO LE), and immobilized on agarose (deGlycIT).
Product Information
IgGZERO (EndoS) is an endoglycosidase from Streptococcus pyogenes that specifically hydrolyzes the complex 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.
IgGZERO 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
- Specific – cleaves only Fc glycans on IgG
- Rapid deglycosylation – 30 min protocol
- Works in serum
- Reduces sample complexity
- Deglycosylates native IgG, no need for denaturation
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
IgGZERO enzyme supplied as lyophilized powder. The enzyme is available in 1000 and 5000 unit formats (1 unit digest 1 µg IgG).
IgGZERO® LE
IgGZERO 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 IgGZERO® - deGlycIT™
deGlycIT is IgGZERO enzyme immobilized on agarose allowing for antibody Fc deglycosylation without contaminating the final preparation with IgGZERO enzyme.
Enzyme Reference Tool
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
- Subunit mass analysis for monitoring antibody oxidation
2017-01-20
Izabela Sokolowska, Jingjie Mo, Jia Dong, Michael J. Lewis, Ping Hu - Calculated conjugated payload from immunoassay and LC-MS intact protein analysis measurements of antibody-drug conjugate
2016-09-29
B. Rago, T. Clark, L. King, J. Zhang, L. N. Tumey, F. Li, F. Barletta, C. Wei, M. Leal,S. Hansel & X. Han
FAQ
Another explanation for not complete deglycosylation could be if the antibody has higher amounts of high-mannose, some hybrid glycans or bisected glycans. For these glycans IgGZERO has low or no activity. Especially, high-mannose glycans are particular resistant to digestion.
If longer incubation time and increased temperature does not increase deglycsylation we would recommend you to try our other endoglycosidase GlycINATOR (EndoS2). This enzyme has much higher activity on these types of glycans sometimes seen in m.Abs.
Yes, many buffers are compatible with IgGZERO/DeGlycIT. Please keep pH close to neutral.
Product Documents
Patent and Disclaimer
IgGZERO®
The trademark IgGZERO is the property of Genovis AB.
For research use only. Not intended for any animal or human therapeutic diagnostic use.