Process of modulating man5 and/or afucosylation content of a glycoprotein composition

Patent metadata

Field	Value
Publication number	US-10059770-B2
Application number	US-201314374163-A
Country	US
Kind code	B2
Filing date	Jan 18, 2013
Priority date	Jan 30, 2012
Publication date	Aug 28, 2018
Grant date	Aug 28, 2018

Abstract

Provided is a method for producing glycoprotein composition with an increased percentage of Man5 and/or afucosylated glycans. Use of manganese for increasing the percentage of Man5 and afucosylated glycans in glycoprotein composition is further provided.

First claim

We claim:

1. A cell culture process for obtaining an antibody composition comprising increased percentage of Man5 glycans and afucosylated glycans by culturing Chinese hamster ovary (CHO) cells expressing said antibody composition in culture medium comprising
a) divalent manganese ion or its salts thereof at a concentration of 2 μM to 20 μM
b) at a pH of about 6.8 to about 7.2
c) recovering the antibody composition from the cell culture.

2. The process according to claim 1 , wherein the antibody composition comprises about 4.5% to about 11% afucosylated glycans.

3. The process according to claim 1 , wherein the antibody composition comprises about 8% to about 22% Man5 glycans.

4. The process according to claim 1 , wherein the antibody composition comprises about 4.5% to about 11% afucosylated glycans and about 8% to about 22% Man5 glycans.

5. The process according to claim 1 , wherein the said process further comprises addition of a feed.

6. The process according to claim 1 , wherein the said process further comprises subjecting cells to a temperature shift wherein cells are cultured a) at first temperature for a first period of time and b) at second temperature for a second period of time.

7. A cell culture process for obtaining an antibody composition comprising increased percentage of Man5 glycans and afucosylated glycans by culturing CHO cells expressing said antibody composition in culture medium comprising the steps of:
a) culturing the cells at a first temperature for a first period of time and a pH of about 6.8 to 7.2;
b) supplementing the cells with divalent manganese ions or its salts thereof at a concentration of 2 μM to 20 μM;
c) subjecting the cells to a temperature shift wherein the cells are cultured at a second temperature for a second period of time; and
d) recovering the antibody composition from the cell culture.

8. The process according to claim 7 , wherein the cells are first cultured at a first temperature of about 37° C.

9. The process according to claim 7 , wherein the cells are shifted to a second temperature of about 31° C. to about 35° C.

10. The process according to claim 7 , wherein the cells are shifted to a second temperature of about 35° C.

11. The process according to claim 7 , wherein the cells are shifted to a second temperature of about 33° C.

12. The process according to claim 7 , wherein the cells are shifted to a second temperature of about 31° C.

13. A cell culture process for obtaining an antibody composition comprising about 8% to about 22% of Man5 glycans and about 4.5% to about 11% of afucosylated glycans by culturing CHO cells expressing said antibody composition in culture medium comprising the steps of:
a) culturing the cells at a first temperature of about 37° C. for a period of time and a pH of about 6.8 to 7.2:
b) supplementing the cells with divalent manganese ions or its salts thereof at a concentration of 2 μM to 20 μM;
c) subjecting the cells to a temperature shift and culturing the cells at a second temperature for a second period of time; and
d) recovering the antibody composition from the cell culture.

Assignees

• Dr Reddys Laboratories Ltd

Inventors

• Thuduppathy Guruvasuthevan Ramasamy
• Madhava Ram Paranandi Ananta
• Bhavsar Kaumil
• Garg Neha

Classifications

• C07K2317/41

  Glycosylation, sialylation, or fucosylation · CPC title

• C12N2500/20

  Transition metals · CPC title

• C12P21/005

  Glycopeptides, glycoproteins · CPC title

• C12N2500/34

  Sugars · CPC title

• C12N1/38

  Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound (C12N1/34 takes precedence) · CPC title