GlycINATOR LE (EndoS2) is an IgG-specific endoglycosidase that rapidly deglycosylates IgG Fc glycans after the core GlcNAc. The GlycINATOR enzyme hydrolyzes all Fc glycoforms on IgG from several species and subclasses. GlycINATOR LE contains low endotoxin levels <0.4 EU/vial and is available as lyophilized powder for deglycosylation of 2 mg of IgG.
| Product Group | |
|---|---|
| Source/Origin | Cloned from S. pyogenes, Produced in E. coli (His-tagged), 92 kDa |
| Product Composition | Lyophilized in TBS, Purified to ≥ 95% homogeneity, Contains low endotoxin levels <0.2 EU/vial |
| Digestion Site | |
| Specificity | Hydrolyzes Beta-1,4-linked GlcNAc residues on all IgG Fc-glycoforms |
| 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 GlycINATOR enzyme binds specifically to the Fc region and therefore will not deglycosylate N-linked glycans in the Fab region.
We can only guarantee optimal deglycosylation for one-time use. We do not have a cleaning or regeneration protocol to provide. However, depending on the antibody and the following application the column can be reused. We do not recommend using with different antibodies due to the risk for contamination of carry-over from previous sample. If column reuse is desired store the column in 10-20% ethanol at +4-8°C.
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.
