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Rhinogen ^® Protein Deglycosylation KitⅡ(for N-Linked & Simple O -Linked Glycans) contains the enzymes and reagents required to remove all N-linked and relatively simple O -linked glycans. The enzyme reagents are a mixture of three glycosidases, including PNGase F, O -Glycosidase, and α2-3.6.8.9Neuraminidase. All enzymes and reagents contained in the kit are compatible with downstream HPLC and MS. PNGase F is a glycosidase that specifically removes N-linked glycans from glycoproteins. It can release most N-glycans (except the innermost GlcNac with α-1,3 fucose residues, which are common in plant and insect glycoproteins) intact from the protein molecules to which they are connected. It can cut between the innermost N-acetylglucosamine (GlcNAc ) and asparagine residues ( Asn ) of the high mannose, hybrid and complex oligosaccharide parts of N-linked glycopeptides or glycoproteins to release complete oligosaccharide chains, as shown in Figure 1.

Figure 1. PNGase F can cleave and release almost all types of N- linked glycans from glycoproteins or glycopeptides.

^, for O -linked glycans, since there is no "universal" endoglycosidase like PNGase F, monosaccharides must be removed by a series of glycosidases. O -Glycosidase and α2-3,6,8,9 Neuraminidase are used to release O - linked sugar chains modified with sialic acid from glycoproteins . Rhinogen ^® α2-3,6,8,9 Neuraminidase can release the non-reducing terminal α (2,3)-, α (2,6)-, α (2,8)-, α (2,9)-sialic acid residues of O - linked sugar chains . After removing the terminal sialic acid residue modification, Rhinogen ^® O -Glycosidase can release the O -linked disaccharides Core 1 (Gal-β(1-3)-GalNAc-) and Core 3 ( GlcNAc -β(1-3)-GalNAc-) linked to the hydroxyl groups of Ser or Thr residues in glycoproteins, as shown in Figure 2.