IgGZERO enzyme is supplied as a lyophilized powder with no preservatives added. It is available in two different sizes; 1000 and 5000 units, for deglycosylation of up to 1 and 5 mg IgG, respectively.
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.
IgGZERO hydrolyzes the β1,4 linkage between the core GlcNAc residues in the Fc-glycan, leaving the innermost GlcNAc intact on the Fc. It is derived from Streptococcus pyogenes and expressed in E. coli. The enzyme contains a His-tag and the molecular weight is 110 kDa.
IgGZERO is specific for N-glycans attached on the Fc-domain of IgGs, and hydrolyzes Fc-glycans of all human IgG subclasses and IgG from many other species, including mouse, rat, monkey, sheep, goat, cow, and horse. The reaction with IgGZERO is mild and runs fast under physiological conditions, with no need for co-factors or detergents.
The IgGZERO enzyme has limited activity on high-mannose and hybrid-type glycans. If you are interested in endoglycosidases that hydrolyze all IgG glycoforms, we recommend the GlycINATOR product family.
Storage and stability
The IgGZERO enzyme is supplied lyophilized and reconstituted by addition of water. After reconstitution, IgGZERO 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.
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.
FAQ and Support
First you could increase incubation time and increase temperature to 37°C.
Another explanation for not complete deglycosylation could be if the antibody has higher amounts of high-mannose, some hybrid glycans or bisected glycans. For these glycans IgGZERO has low or no activity. Especially, high-mannose glycans are particular resistant to digestion.
If longer incubation time and increased temperature does not increase deglycsylation we would recommend you to try our other endoglycosidase GlycINATOR (EndoS2). This enzyme has much higher activity on these types of glycans sometimes seen in m.Abs.
Patent and Disclaimer
The trademark IgGZERO is the property of Genovis AB.
For research use only. Not intended for any animal or human therapeutic diagnostic use.