SmartEnzymes IgGZERO workflow

Hydrolysis of Fc N-glycans

IgGZERO (EndoS) is an IgG-specific endoglycosidase hydrolyzing complex N-glycans at the Fc N-glycosylation sites.

The enzyme 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.
 

  • 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
Hydrolyzes the β1,4-linkage between the two innermost GlcNAc residues. Limited activity on high-mannose and hybrid-type Fc N-glycans
30 min reaction
Requires native IgG fold

IgGZERO® Lyophilized

Lyophilized enzyme for deglycosylation of IgG

IgGZERO® Low Endotoxin

Lyophilized enzyme with low levels of endotoxin for deglycosylation of IgG

IgGZERO® Immobilized

Immobilized enzyme for deglycosylation of IgG in spin columns

About IgGZERO®

IgGZERO workflow

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 (Sjögren et al., 2015). If you are interested in endoglycosidases that hydrolyze all Fc N-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.

IgGZERO® Lyophilized

Lyophilized enzyme for deglycosylation of IgG.
IgGZERO Lyophilized is available as a lyophilized powder in two different sizes; 1000 and 5000 units, for deglycosylation of 1 or 5 mg IgG, respectively.
 

  • Flexible format for method development
  • Possible to combine enzymes for improved efficiency
  • Amenable to automated workflows
  • Ready to use – just add water!

Available Products

IgGZERO Lyophilized 1000 unitsLyophilized enzyme for deglycosylation of 1 mg IgGA0-IZ1-010309.00Buy / Request a Quote
IgGZERO Lyophilized 5000 unitsLyophilized enzyme for deglycosylation of 5 mg IgGA0-IZ1-0501,158.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One unit deglycosylates ≥ 95% of 1μg human IgG, when incubated in 10 mM sodium phosphate, 150 mM NaCl, pH 7.4 at 37°C for 30 min.

Content and Storage

IgGZERO Lyophilized is reconstituted by addition of water. No preservatives added. After reconstitution, IgGZERO Lyophilized is stable for 1 month at +4-8°C. The product is shipped at ambient temperature and should be stored at -20°C upon arrival.

IgGZERO® Low Endotoxin

Lyophilized enzyme with low levels of endotoxin for deglycosylation of IgG.

IgGZERO Low Endotoxin is available as a lyophilized powder in 2000 unit vials for deglycosylation of 2 mg IgG. This product formulation has low levels of endotoxins per vial (< 0.2 EU).

 

  • Efficient digestion with documented low levels of endotoxin
  • Suitable for use in sensitive in vivo or cell-based assays

Available Products

IgGZERO Low Endotoxin 2000 unitsLyophilized enzyme with low levels of endotoxin for deglycosylation of 2 mg IgGA0-IZ8-020639.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One unit deglycosylates ≥ 95% of 1μg human IgG when incubated in 10 mM sodium phosphate, 150 mM NaCl, pH 7.4 at 37°C for 30 min.

Content and Storage

IgGZERO Low Endotoxin is reconstituted by addition of water. No preservatives added. After reconstitution, IgGZERO Low Endotoxin is stable for 1 month at +4-8°C. The product is shipped at ambient temperature, and should be stored at -20°C upon arrival.

Instructions

Product Specification

Safety Data Sheet

Certificate of Analysis

IgGZERO® Immobilized (formerly known as deGlycIT)

Immobilized enzyme for deglycosylation of IgG in spin columns.

The IgGZERO Immobilized spin columns contain the IgGZERO enzyme covalently coupled to agarose beads, for deglycosylation of Fc N-glycans without contaminating the final preparation with enzyme.

The spin columns are available for deglycosylation of 0.5 mg (Microspin), 10 mg (Midispin) or 100 mg (Maxispin) IgG. The smallest columns are available in three different pack sizes, with 2, 5 or 10 columns per pack.

 

  • Easy-to-use spin columns
  • Increased enzyme-to-substrate ratio for improved digestion efficiency
  • No enzyme in the final sample

Available Products

IgGZERO Immobilized Microspin 2 x 0.5 mgImmobilized enzyme for deglycosylation of 2 x 0.5 mg IgG in spin columnsA0-IZ6-010430.00Buy / Request a Quote
IgGZERO Immobilized Microspin 5 x 0.5 mgImmobilized enzyme for deglycosylation of 5 x 0.5 mg IgG in spin columnsA0-IZ6-025970.00Buy / Request a Quote
IgGZERO Immobilized Microspin 10 x 0.5 mgImmobilized enzyme for deglycosylation of 10 x 0.5 mg IgG in spin columnsA0-IZ6-0501,599.00Buy / Request a Quote
IgGZERO Immobilized Midispin 1-10 mgImmobilized enzyme for deglycosylation of 1-10 mg IgG in spin columnsA0-IZ6-1001,290.00Buy / Request a Quote
IgGZERO Immobilized Maxispin 10-100 mgImmobilized enzyme for deglycosylation of 10-100 mg IgG in spin columnsA0-IZ6-10003,847.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One spin column contains sufficient IgGZERO-coupled agaose beads to deglycosylate 0.5 mg (Microspin), 10 (Midispin) or 100 (Maxispin) mg IgG.

Content and Storage

IgGZERO Immobilized is supplied in 20% ethanol, shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the product!

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.

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.

 

Find all FAQ here

Order