Core fucosylated glycopeptides and glycoproteins: chemoenzymatic synthesis and uses thereof

A method of preparing a core-fucosylated glycopeptide or core-fucosylated glycoprotein having a predetermined oligosaccharide moiety, the method comprising:

• providing a core-fucosylated acceptor glycopeptide or core-fucosylated acceptor glycoprotein comprising an asparagine-linked N-acetylglucosamine (GlcNAc) residue linked to a core fucose residue, wherein the core-fucosylated acceptor glycopeptide or core-fucosylated acceptor glycoprotein is an alpha-1-6-fucosyl-GlcNAc-peptide or an alpha-1-6-fucosyl-GlcNAc-protein;
• and
• enzymatically reacting the core-fucosylated acceptor glycoprotein or core-fucosylated acceptor glycopeptide with an activated oligosaccharide donor in the presence of a buffer and at least one fusion protein comprising a cysteine protease domain (CPD) and an endoglycosidase selected from the group consisting of Endoglycosidase-F2 D124A mutant (SEQ ID NO:
• 4), Endoglycosidase-F2 D124Q mutant (SEQ ID NO:
• 5), Endoglycosidase-F3 D126A mutant (SEQ ID NO:
• 6), and Endoglycosidase-F3 D126Q mutant (SEQ ID NO:
• 7), wherein the activated oligosaccharide donor comprises an oxazoline covalently linked to a oligosaccharide moiety, wherein the oligosaccharide moiety comprises a predetermined number and type of sugar residues, and wherein the enzymatic reaction covalently links the oligosaccharide moiety to the core-fucosylated acceptor glycoprotein or core-fucosylated acceptor glycopeptide;
• thereby providing the core-fucosylated glycoprotein or core-fucosylated acceptor glycopeptide having the predetermined oligosaccharide moiety.