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IgGZERO™ – Hydrolysis of Complex-type Fc N-glycans
IgGZERO (EndoS) is an IgG-specific endoglycosidase hydrolyzing complex N-glycans at the Fc N-glycosylation sites.
SmartEnzymes™
IgGZERO deglycosylates IgG after the core GlcNAc and display limited activity on high-mannose and hybrid-type glycans. The enzyme is used to rapidly reduce sample complexity, to inactivate Fc-mediated effector functions in immunoassays and as a tool to improve imaging by reducing Fc interactions.
Why IgGZERO™?
- Highly efficient IgG-specific endoglycosidase
- Fast – hydrolyzes complex Fc N-glycoforms within 30 minutes
- Enables characterization and quality control of antibody-based biotherapeutics
- Available immobilized in ready-to-use spin columns
Human IgG1-4, Fc-fusion proteins, IgG from mouse, rabbit, rat, monkey, sheep, goat, cow and horse
30 min reaction
Requires native IgG fold
Hydrolyzes the β1,4-linkage between the two innermost GlcNAc residues. Limited activity on high-mannose and hybrid-type Fc N-glycans
Product Formats
IgGZERO™ Immobilized
Immobilized enzyme for hydrolysis of complex-type Fc N-glycans in spin columns
Buy productIgGZERO™ Low Endotoxin
Lyophilized enzyme with low levels of endotoxin for hydrolysis of complex-type Fc N-glycans
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About IgGZERO™
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 IgG 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. If you are interested in endoglycosidases that hydrolyze all Fc N-glycoforms on IgG, we recommend the GlycINATOR product family.
Applications
A simple and rapid estimation of IgG core fucosylation using LC-MS analysis.
Citations
FAQ and Support
Can I use PBS 1X instead of the IgGZERO reaction buffer?
Yes, the IgGZERO enzyme is 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.
I read that cleavage buffer for IgGZERO is 10 mM sodium phosphate, 150 mM NaCl, pH 7.4 Can I use PBS 1X in place of cleavage buffer?
Yes, many buffers are compatible with IgGZERO. Please keep pH close to neutral.
Which should I choose, GlycINATOR or IgGZERO?
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.
What is the difference between GlycINATOR and IgGZERO?
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.
I did not get complete deglycosylation using IgGZERO Immobilized, how can I improve digestion?
First you could increase incubation time and increase temperature to 37°C. One explanation for incomplete deglycosylation with IgGZERO Immobilized 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.
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