SmartEnzymes™

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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:

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.

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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.

Immobilized IgGZERO® - deGlycIT™

deGlycIT is IgGZERO enzyme immobilized on agarose allowing for antibody Fc deglycosylation without contaminating the final preparation with IgGZERO enzyme.
 


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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.

In both products the enzyme is immobilized on agarose beads for IgG-specific hydrolysis of the Fc glycans to the innermost GlcNAc. The deGlycIT contains immobilized IgGZERO (EndoS) and Immobilized GlycINATOR contains GlycINATOR (EndoS2). The GlycINATOR enzyme has much higher enzymatic activity for high mannose and some bisected and hybrid glycans that can occur on mAbs.

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.

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.

Yes, many buffers are compatible with IgGZERO/DeGlycIT. Please keep pH close to neutral.

 

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