Cell culture process
DC CAFC
First Claim
1. A process for producing a human glycoprotein having multiple glycoforms, comprising culturing Chinese hamster ovary cells expressing nucleic acid encoding said glycoprotein in a serum-free medium in a production phase at a temperature of about 30°
- C. to 35°
C. and in the presence of about 0 to 2 mM of a butyrate salt, wherein the process produces an increased percentage of glycoprotein molecules having one glycoform relative to an identical process performed at 37°
C. in the absence of butyrate.
0 Assignments
Litigations
0 Petitions
Accused Products
Abstract
A glycoprotein is produced by a process comprising culturing mammalian host cells expressing nucleic acid encoding a glycoprotein in the presence of (a) a factor that modifies growth state in a cell culture, (b) a divalent metal cation that can adopt and prefers an octahedral coordination geometry, and/or (c) a plasma component. In this process, the occupancy of an N-linked glycosylation site occupied only in a fraction of a glycoprotein is enhanced. Such culturing is preferably carried out at a temperature of between about 30° C. and 35° C. and/or in the presence of up to about 2 mM of a butyrate salt and/or in the presence of a cell-cycle inhibitor.
46 Citations
1 Claim
-
1. A process for producing a human glycoprotein having multiple glycoforms, comprising culturing Chinese hamster ovary cells expressing nucleic acid encoding said glycoprotein in a serum-free medium in a production phase at a temperature of about 30°
- C. to 35°
C. and in the presence of about 0 to 2 mM of a butyrate salt, wherein the process produces an increased percentage of glycoprotein molecules having one glycoform relative to an identical process performed at 37°
C. in the absence of butyrate.
- C. to 35°
Specification
-
- Resources
-
Current AssigneeGenentech, Inc. (Roche Holding AG)
-
Original AssigneeGenentech, Inc. (Roche Holding AG)
-
InventorsBridges, Tiffany M., Gawlitzek, Martin, Andersen, Dana C., Hoy, Cynthia A.
-
Primary Examiner(s)Carlson, Karen Cochrane
-
Assistant Examiner(s)Mitra, Rita
-
Application NumberUS09/723,545Time in Patent Office1,002 DaysField of Search530/395, 435/69.1, 435/69.6, 435/252.3, 435/358, 435/359, 435/70.1US Class Current435/70.1CPC Class CodesC12N 9/6459 t-plasminogen activator (3....C12P 21/005 Glycopeptides, glycoproteinsC12Y 304/21069 Protein C activated (3.4.21...
