Methods for controlling the galactosylation profile of recombinantly-expressed proteins

Patent metadata

Field	Value
Publication number	US-2016152702-A1
Application number	US-201615014694-A
Country	US
Kind code	A1
Filing date	Feb 3, 2016
Priority date	Apr 27, 2011
Publication date	Jun 2, 2016
Grant date	—

Abstract

The present invention relates to methods for modulating the glycosylation profile of recombinantly-expressed proteins. In particular, the present invention relates to methods of controlling the galactosylation profile of recombinantly-expressed proteins by supplementing production medium, e.g., a hydrolysate-based or a chemically defined medium, with manganese and/or D-galactose

First claim

What is claimed is:

1 . A composition comprising an antibody comprising the heavy and light chain variable domains of adalimumab, wherein less than 70% of the total N-linked oligosaccharides present on said antibody are of an agalactosyl fucosylated biantennary oligosaccharide form (sum NGA2F+NGA2F-GlcNAc).

2 . The composition of claim 1 , wherein the antibody is adalimumab.

3 . The composition of claim 1 , wherein 49-69% of the total N-linked oligosaccharides present on said antibody are of an agalactosyl fucosylated biantennary oligosaccharide form (sum NGA2F+NGA2F-GlcNAc).

4 . The composition of claim 1 , wherein 55-65% of the total N-linked oligosaccharides present on said antibody are of an agalactosyl fucosylated biantennary oligosaccharide form (sum NGA2F+NGA2F-GlcNAc).

5 . The composition of claim 1 , wherein 26-40% of the total N-linked oligosaccharides present on said antibody are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F).

6 . The composition of claim 1 , wherein 35-44% of the total N-linked oligosaccharides present on said antibody are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F).

7 . The composition of claim 1 , wherein 30-40% of the total N-linked oligosaccharides present on said antibody are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F).

8 . The composition of claim 1 , wherein said antibody has been produced by culturing a mammalian cell in a cell culture media which has been supplemented with a manganese supplement and a galactose supplement.

9 . The composition of claim 8 , wherein the cell culture media has been supplemented with 0.2-100 μM of a manganese supplement and 1-100 mM of a galactose supplement.

10 . The composition of claim 8 , wherein the cell culture media comprises a hydrolysate based media or a chemically defined media.

11 . The composition of claim 8 , wherein the mammalian cell is a CHO cell or an NSO cell.

12 . A pharmaceutical composition comprising the composition of claim 1 and a pharmaceutically acceptable carrier.

13 . The pharmaceutical composition of claim 12 , wherein the pharmaceutically acceptable carrier comprises a polyalcohol.

14 . The pharmaceutical composition of claim 12 , wherein the pharmaceutically acceptable carrier comprises mannitol, sodium chloride, sodium phosphate or polysorbate 80.

15 . The pharmaceutical composition of claim 12 , wherein the pharmaceutical composition is suitable for parenteral administration.

16 . A composition comprising adalimumab, wherein less than 70% of the total N-linked oligosaccharides present on said adalimumab are of an agalactosyl fucosylated biantennary oligosaccharide form (sum NGA2F+NGA2F-GlcNAc).

17 . The composition of claim 16 , wherein 49-69% of the total N-linked oligosaccharides present on said adalimumab are of an agalactosyl fucosylated biantennary oligosaccharide form (sum NGA2F+NGA2F-GlcNAc).

18 . The composition of claim 16 , wherein 55-65% of the total N-linked oligosaccharides present on said adalimumab are of an agalactosyl fucosylated biantennary oligosaccharide form (sum NGA2F+NGA2F-GlcNAc).

19 . The composition of claim 16 , wherein 26-40% of the total N-linked oligosaccharides present on said adalimumab are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F).

20 . The composition of claim 16 , wherein 35-44% of the total N-linked oligosaccharides present on said adalimumab are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F).

21 . The composition of claim 16 , wherein 30-40% of the total N-linked oligosaccharides present on said adalimumab are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F).

22 . The composition of claim 16 , wherein said adalimumab has been produced by culturing a mammalian cell in a cell culture media which has been supplemented with a manganese supplement and a galactose supplement.

23 . The composition of claim 22 , wherein the cell culture media has been supplemented with 0.2-100 μM of a manganese supplement and 1-100 mM of a galactose supplement.

24 . The composition of claim 22 , wherein the cell culture media comprises a hydrolysate based media or a chemically defined media.

25 . The composition of claim 22 , wherein the mammalian cell is a CHO cell or an NSO cell.

26 . A pharmaceutical composition comprising the composition of claim 16 and a pharmaceutically acceptable carrier.

27 . The pharmaceutical composition of claim 26 , wherein the pharmaceutically acceptable carrier comprises a polyalcohol.

28 . The pharmaceutical composition of claim 26 , wherein the pharmaceutically acceptable carrier comprises mannitol, sodium chloride, sodium phosphate or polysorbate 80.

29 . The pharmaceutical composition of claim 26 , wherein the pharmaceutical composition further comprises polysorbate 80.

30 . The pharmaceutical composition of claim 26 , wherein the pharmaceutical composition is suitable for parenteral administration.

Assignees

• Abbvie Inc

Inventors

• Bengea Cornelia
• Rives Lisa M
• Hossler Patrick

Classifications

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title

• C07K2317/41

  Glycosylation, sialylation, or fucosylation · CPC title

• C12N2500/34

  Sugars · CPC title

• C07K2317/21

  from primates, e.g. man · CPC title

• C07K2317/14

  Specific host cells or culture conditions, e.g. components, pH or temperature · CPC title