John L Giddens

US Patent:

20120226024, Sep 6, 2012

Filed:

Mar 5, 2012

Appl. No.:

13/411733

Inventors:

LAI-XI WANG - ELLICOTT CITY MD, US
WEI HUANG - DUNDALK MD, US
JOHN GIDDENS - CHESAPEAKE VA, US

International Classification:

C12P 21/06
C07K 16/00
C12P 21/08

US Classification:

5303873, 435 681

Abstract:

A chemoenzymatic method for the preparation of a core-fucoslyated glycoprotein or glycopeptide, including (a) providing an acceptor selected from the group consisting of a fucosylated GlcNAc-protein and fucosylated GlcNAc-peptide; and (b) reacting the acceptor with a donor substrate including an activated oligosaccharide moiety, in the presence of an endoglycosidase (ENGase) selected from Endo;F1, Endo-F2, Endo-F3, Endo-D and related glycosynthase mutants to transfer the oligosaccharide moiety to the acceptor and yield the structure defined core-fucosylated glycoprotein or glycopeptide. The donor substrate includes, in a specific implementation, a synthetic oligosaccharide oxazoline. A related method of fucosylated glycoprotein or fucosylated glycopeptide remodeling with a predetermined natural N-glycan or a tailor-made oligosaccharide moiety, and a method of remodeling an antibody to include a predetermined sugar chain to replace a heterogeneous sugar chain, are also described.