Patent ID: 9175326
Filing Date: 2015-11-03
Classification: A61K,C07K,C12N,C12P,C12Y

Abstract:
1. A method of preparing a fucosylated or nonfucosylated glycoprotein having a predetermined oligosaccharide moiety, the method comprising: providing a fucosylated or nonfucosylated GlcNAc acceptor protein comprising an asparagine-linked N-acetylglucosamine (GlcNAc) residue, wherein the fucosylated acceptor protein is an alpha-1-6-fucosyl-GlcNAc-protein and the nonfucosylated acceptor protein is an alpha-1-6-GlcNAc-protein; and enzymatically reacting the fucosylated or nonfucosylated GlcNAc acceptor protein with an activated oligosaccharide donor in the presence of an Endoglycosidase-D full length or truncated mutant selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO: 9, wherein the activated oligosaccharide donor comprises an oxazoline covalently linked to an oligosaccharide moiety, wherein the oligosaccharide moiety comprises a mannose type N-glycan having a predetermined number of mannose residues, wherein the enzymatically reacting covalently links the oligosaccharide moiety to the fucosylated or nonfucosylated GlcNAc acceptor protein, thereby providing the fucosylated or nonfucosylated glycoprotein having the predetermined oligosaccharide moiety.