IP Library › Granted Patent US 12,209,261
Granted Patent B2
US 12,209,261 · App. 15/780,344 · Granted Jan 28, 2025

Media for the expression of recombinant vitamin K-dependent proteins

Inventors: Yih Yean Lee (Victoria, AU); Chee Kin Andrew Low (Victoria, AU); Campbell Douglas Aitken (Victoria, AU); Steven Patrick Byrne (Victoria, AU); Mark John Ferres Edwards (Victoria, AU)
Assignee: CSL BEHRING LENGNAU AG
C12N9/6437C07K14/4702C07K14/745C07K14/76C12N5/0682C12N5/16A61K38/00C07K2319/31C12N2500/30C12N2500/44
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Quick Facts
Patent No.
US 12,209,261
App. No.
15/780,344
Granted
Jan 28, 2025
Kind
B2
Abstract

The present invention relates to a method for increasing the activity and/or the yield of a recombinant vitamin K-dependent protein expressed in cell culture. The present invention further relates to uses and compositions of matter.

Claims (19)

1. A method comprising:

culturing mammalian host cells that comprise an expression system for expressing a recombinant vitamin K-dependent protein under conditions suitable for achieving expression of the recombinant vitamin K-dependent protein in a cell culture medium comprising one or more cell culture enhancing reagent(s) at concentration(s) sufficient to enhance the activity of the recombinant vitamin K-dependent protein compared to a recombinant vitamin K-dependent protein expressed in mammalian host cells cultured in a cell culture medium lacking the cell culture enhancing reagent(s),

wherein the one or more cell culture enhancing reagent(s) and concentration(s) is/are chosen from

alpha-ketoglutaric acid at a concentration of 7.5-60 mmol/L,

succinic acid at a concentration of 1.8-30 mmol/L,

oxaloacetic acid at a concentration of 7.5-30 mmol/L,

malic acid at a concentration of 7.5-30 mmol/L,

fumaric acid at a concentration of 3.7-30 mmol/L, and

citric acid at a concentration of 0.9-15 mmol/L.

2. The method of claim 1 , wherein the cell culture medium comprises a combination of any two or more of alpha-ketoglutaric acid, succinic acid, oxaloacetic acid, malic acid, fumaric acid, and citric acid.

3. The method of claim 1 , comprising providing the one or more cell culture enhancing reagent(s) at the beginning of the cell culturing.

4. The method of claim 1 , wherein the cell culture is a fed-batch culture, and wherein the one or more cell culture enhancing reagent(s) is/are present in a basal cell culture medium and/or in a feed medium.

5. The method of claim 1 , wherein the cell culture is a perfusion culture, and wherein the one or more cell culture enhancing reagent(s) is/are present in a basal cell culture medium and/or in a perfusion medium.

6. The method of claim 1 , wherein the cell culture medium is free of any proteins from human or animal origin.

7. The method of claim 1 , wherein the host cells are Chinese hamster ovary (CHO) cells.

8. The method of claim 1 , wherein the recombinant vitamin K-dependent protein is selected from Factor IX (FIX), Factor VII (FVII), Factor X (FX), Factor II (FII), Protein C, Protein S, Protein Z, osteocalcin, the calcification inhibiting matrix Gla protein (MGP), and the cell growth regulating growth arrest specific protein 6 (Gas6).

9. The method of claim 1 , wherein the recombinant vitamin K-dependent protein is a FVII protein.

10. The method of claim 1 , wherein the recombinant vitamin K-dependent protein is a FVII fusion protein.

11. The method of claim 1 , wherein the recombinant vitamin K-dependent protein is a FVII albumin fusion protein.

Assignments (4)
ASSIGNEE CHANGE OF ADDRESS Recorded Mar 12, 2021
From: CSL BEHRING LENGNAU AG
To: CSL BEHRING LENGNAU AG
Reel/Frame 055580/0102 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 8, 2018
From: LEE, YIH YEAN; LOW, CHEE KIN ANDREW; AITKEN, CAMPBELL DOUGLAS; BYRNE, STEVEN; EDWARDS, MARK JOHN FERRES
To: CSL LIMITED
Reel/Frame 047448/0505 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 8, 2018
From: CSL LIMITED
To: CSL BEHRING RECOMBINANT FACILITY AG
Reel/Frame 047448/0603 →
CHANGE OF NAME Recorded Nov 8, 2018
From: CSL BEHRING RECOMBINANT FACILITY AG
To: CSL BEHRING LENGNAU AG
Reel/Frame 047450/0544 →
Priority Claims (1)
EP 15197516 · Dec 2, 2015 · regional
Continuity (1)
Related Publication 20180362952A1 · Dec 20, 2018
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