Methods for controlling the galactosylation profile of recombinantly-expressed proteins

Patent metadata

Field	Value
Publication number	US-9255143-B2
Application number	US-201514619799-A
Country	US
Kind code	B2
Filing date	Feb 11, 2015
Priority date	Apr 27, 2011
Publication date	Feb 9, 2016
Grant date	Feb 9, 2016

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 adalimumab, wherein more than 25% of the total N-linked oligosaccharides present on said adalimumab are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F).

2. The composition of claim 1 , 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).

3. The composition of claim 1 , 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).

4. The composition of claim 1 , 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).

5. The composition of claim 1 , 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).

6. The composition of claim 5 , 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).

7. The composition of claim 5 , 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).

8. The composition of claim 1 , 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.

9. The composition of claim 1 , wherein said adalimumab has been produced by culturing a mammalian cell in a cell culture media comprising 0.2-100 μM of a manganese supplement and 1-100 mM of a galactose supplement.

10. The composition of claim 9 , wherein the cell culture media comprises a hydrolysate based media or a chemically defined media.

11. The composition of claim 9 , wherein the mammalian cell is a CHO cell.

12. The pharmaceutical composition of claim 11 and a pharmaceutically acceptable carrier comprising a polyalcohol.

13. The pharmaceutical composition of claim 12 , wherein the pharmaceutically acceptable carrier further comprises polysorbate 80.

14. The pharmaceutical composition of claim 13 , wherein the pharmaceutical composition is suitable for subcutaneous administration and is formulated in a pre-filled syringe.

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

16. The pharmaceutical composition of claim 15 , wherein the pharmaceutically acceptable carrier comprises mannitol.

17. The pharmaceutical composition of claim 15 , wherein the pharmaceutically acceptable carrier comprises sodium chloride.

18. The pharmaceutical composition of claim 15 , wherein the pharmaceutically acceptable carrier comprises polysorbate 80.

19. The pharmaceutical composition of claim 15 , wherein the pharmaceutically acceptable carrier comprises sodium phosphate.

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

21. The pharmaceutical composition of claim 15 , wherein the pharmaceutical composition is suitable for subcutaneous administration and is formulated in a pre-filled syringe.

Assignees

• Abbvie Inc
• Abbvie Inc

Inventors

• Bengea Cornelia
• Rives Lisa M
• Hossler Patrick

Classifications

• C07K2317/14

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

• C07K16/00

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

• C12N2500/34

  Sugars · CPC title

• C07K16/241 Primary

  Tumor Necrosis Factors · CPC title

• C12N5/0018

  Culture media for cell or tissue culture (media for specific animal cell type C12N5/06) · CPC title