IgGZERO LE (EndoS) is an IgG-specific endoglycosidase that rapidly deglycosylates biantennary Fc glycans after the core GlcNAc on IgG from several species and subclasses. IgGZERO LE contains low endotoxin levels <0.4 EU/vial and is available as lyophilized powder for deglycosylation of 1-5 mg of IgG.
| Product Group | |
|---|---|
| Source/Origin | Cloned from S. pyogenes, Produced in E. coli (His-tagged), 112 kDa |
| Product Composition | Lyophilized in PBS, Purified to ≥ 95% homogeneity, Contains low endotoxin levels <0.2 EU/vial |
| Digestion Site | |
| Specificity | |
| Substrates | Cow IgG, Fc-fusion proteins, Goat IgG, Horse IgG, Human IgG1, Human IgG2, Human IgG3, Human IgG4, Monkey IgG, Mouse IgG, Rabbit IgG, Rat IgG, Sheep IgG |
| Reaction Conditions | 30 minute reaction at 37°C according to protocol specifications |
| Digestion Buffers | |
| Buffer Compatibility | |
| Inhibitors | |
| Unit Definition | |
| Application | Afucosylation Assessment, Antibody Deglycosylation, Immunoassays |
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.
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.
Yes, many buffers are compatible with IgGZERO. Please keep pH close to neutral.
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.
In both products the enzyme is immobilized on agarose beads for IgG-specific hydrolysis of the Fc glycans to the innermost GlcNAc. IgGZERO Immobilized contains IgGZERO (EndoS) and GlycINATOR Immobilized 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.
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.
