Question 1

Is it possible to reuse the OmniGLYZOR column?

Maria Ekemohn · Accepted Answer

No, it is not recommended. We can only guarantee optimal performance for one-time use.

Question 2

Can OmniGLYZOR be used for released glycan analysis?

Maria Ekemohn · Accepted Answer

No, both N- and O-glycans are trimmed by the exoglycosidases present in OmniGLYZOR and do not longer represent the structures found on the intact glycoprotein substrate.

Question 3

Does OmniGLYZOR remove all N-glycans?

Maria Ekemohn · Accepted Answer

OmniGLYZOR hydrolyzes the amide bond between the polypeptide asparagine and the innermost GlcNAc of all mammalian asparagine-linked complex, hybrid, or high mannose oligosaccharides. It does not remove N-glycans with α1-3 core fucosylation.

Question 4

Does OmniGLYZOR remove all N-glycans?

Maria Ekemohn · Accepted Answer

OmniGLYZOR hydrolyzes the amide bond between the polypeptide asparagine and the innermost GlcNAc of all mammalian asparagine-linked complex, hybrid, or high mannose oligosaccharides. It does not remove N-glycans with α1-3 core fucosylation.

Question 5

Can I re-use the GlycINATOR Immobilized column?

Felicia Mejare · Accepted Answer

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.

Question 6

Can I achieve Fc-specific deglycosylation on IgG with N-linked glycans in the Fab region?

Felicia Mejare · Accepted Answer

Yes, the GlycINATOR enzyme binds specifically to the Fc region and therefore will not deglycosylate N-linked glycans in the Fab region.

Question 7

What is the difference between GlycINATOR Immobilized and IgGZERO Immobilized?

Felicia Mejare · Accepted Answer

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.

Question 8

Which should I choose, GlycINATOR or IgGZERO?

Felicia Mejare · Accepted Answer

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.

Question 9

What is the difference between GlycINATOR and IgGZERO?

Felicia Mejare · Accepted Answer

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.

Question 10

What is the difference between GlycINATOR and IgGZERO?

Felicia Mejare · Accepted Answer

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.

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