deGlycIT™ (Immobilized IgGZERO®)
deGlycIT spin columns contain IgGZERO enzyme covalently coupled to agarose beads for deglycosylation of Fc-glycans without contaminating the final preparation with enzyme.
IgGZERO (EndoS) is an IgG-specific endoglycosidase acting on complex type N-glycans at the Fc-glycosylation site of IgG.
Immobilized IgGZERO enzyme (deGlycIT) on agarose in convenient spin columns. The spin columns are available in three different sizes for deglycosylation of up to 0.5 (MicroSpin), 10 (MidiSpin) or 100 (MaxiSpin) mg IgG.
The smallest columns are available in three different pack sizes with 2, 5 or 10 columns per pack. deGlycIT hydrolyzes the β1,4 linkage between the core GlcNAc residues in the Fc-glycan, leaving the innermost GlcNAc intact on the Fc. IgG is incubated with the agarose beads for 15 min, and deglycosylated IgG is then collected by a 1 minute centrifugation step. Since deGlycIT is immobilized on agarose beads, there is no need for extensive purification to remove the enzyme.
deGlycIT deglycosylates IgG under physiological reaction conditions, thus preserving the immunoreactivity. Deglycosylation is performed at room temperature.
The deGlycIT enzyme has limited activity on high-mannose and hybrid-type glycans. If you are interested in endoglycosidases that hydrolyze these types of glycans, we recommend the GlycINATOR product family.
Storage and stability
deGlycIT is supplied in 20% ethanol, shipped on ice, and should be stored at +4-8°C upon arrival. Do not freeze the product!
One spin column contains sufficient IgGZERO-coupled agaose beads to deglycosylate 0.5 mg (MicroSpin), 10 (MidiSpin) or 100 (MaxiSpin) mg IgG.
FAQ and Support
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 deGlycIT is the property of Genovis AB.
For research use only. Not intended for any animal or human therapeutic diagnostic use.