Herein is reported a method for the enzymatic production of an antibody with a modified glycosylation in the Fc-region comprising the steps of incubating the antibody light chain affinity ligand-bound monoclonal antibody with a glycosylation in the Fc-region with a first enzyme for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region, recovering the antibody from the antibody light chain affinity ligand, incubating the recovered antibody in solution with a second enzyme for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region to a defined form, separating the antibody with the modified glycosylation in the Fc-region from the second enzyme in a cation exchange chromatography, and thereby producing the antibody with a modified glycosylation in the Fc-region.

Methods for making modified Fc regions of antibodies and antibody fragments, both human and humanized, and having enhanced stability and efficacy, are provided. Antibodies comprising Fc regions with core fucose residues removed, and attached to oligosaccharides comprising terminal sialyl residues, are provided. Antibodies comprising homogeneous glycosylation of Fc regions with specific oligosaccharides are provided. Fc regions conjugated with homogeneous glycoforms of monosaccharides and trisaccharides, are provided. Methods of preparing human antibodies with modified Fc using glycan engineering, are provided.

Glycoproteins having particular sialylation patterns, and methods of making and using such glycoproteins, are described.

Disclosed herein are methods of generating proteoglycans with distinct glycan structures in engineered, non-naturally occurring eukaryotic cells. These methods make accessible a dynamic range of protein glycosylation. Compositions of engineered, non-naturally occurring cells capable of generating these proteoglycans are also disclosed herein.

Glycoproteins having particular sialylation patterns, and methods of making and using such glycoproteins, are described.

Glycoproteins having particular sialylation patterns, and methods of making and using such glycoproteins, are described.

Herein is reported a method for the enzymatic production of an antibody with a modified glycosylation in the Fc-region comprising the steps of incubating the antibody light chain affinity ligand-bound monoclonal antibody with a glycosylation in the Fc-region with a first enzyme for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region, recovering the antibody from the antibody light chain affinity ligand, incubating the recovered antibody in solution with a second enzyme for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region to a defined form, separating the antibody with the modified glycosylation in the Fc-region from the second enzyme in a cation exchange chromatography, and thereby producing the antibody with a modified glycosylation in the Fc-region.

Glycoproteins having particular sialylation patterns, and methods of making and using such glycoproteins, are described.

This disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, this disclosure is in the technical field of cultivation of metabolically engineered cells. This disclosure describes a method for the production of a glycosylated product derived from UDP-GlcNAc and comprising a di- or oligosaccharide that is composed of at least two different monosaccharide subunits by a cell as well as the separation of the glycosylated product from the cultivation. Furthermore, this disclosure provides a metabolically engineered cell for production of a glycosylated product derived from UDP-GlcNAc and comprising a di- or oligosaccharide that is composed of at least two different monosaccharide subunits. This disclosure also provides a cell excreting a di- or oligosaccharide out of the cell.

Glycoproteins having particular sialylation patterns, and methods of making and using such glycoproteins, are described.