Culture media comprising n-Acyl-X-glutamine dipeptides
Disclosed herein is a cell culture media containing L-glutamine from a set of N-acylated dipeptides Acyl-X-Q, and L-glutamine from a set of other glutamine-sources Qsource in a defined molar ratio R=n(Acyl-X-Q)/n(Qsource), wherein the variables X, Q, Acyl, R, n(Acyl-X-Q) and n(Qsource) are defined in the general disclosure. Processes of using the cell culture media are also described herein.
1. A cell culture media, comprising
L-glutamine from a set of N-acylated dipeptides Acyl-XQ; and
L-glutamine from a set of other glutamine-sources Qsource in a defined molar ratio R=n(Acyl-X-Q)/n(Qsource),
wherein:
X is defined as an L-amino acid;
Q is defined as L-glutamine attached via an amide bond to L-amino acid X;
Acyl is defined as a C1-C7-acyl moiety attached via an amide bond to the amino-terminus of L-amino acid X;
R is defined to be in the range of 0.03 to 20;
n(Acyl-X-Q) is the total amount of substance of L-glutamine contained in the set of N-acylated dipeptides Acyl-X-Q in the culture media;
n(Qsource) is the total amount of substance of L-glutamine contained in the set of other glutamine sources Qsource in the culture media;
the constituents of the set of other L-glutamine sources Qsource are selected from the following: Free L-glutamine, dipeptides Y-Q, or mixtures thereof, wherein Y is defined as one of the 20 genetically encoded L-amino acids, wherein Q is defined as L-glutamine attached via an amide bond to L-amino acid Y, and the amide bond connecting Y and Q in dipeptide Y-Q is a regular backbone amide bond involving the carboxy terminus of amino acid Y and the amino terminus of glutamine Q; and
the cell culture media are serum free.
2. The cell culture media according to claim 1 , wherein the cell culture media are chemically defined.
3. The cell culture media according to claim 1 , wherein the only constituent of the set of other L-glutamine sources Qsource is dipeptide Y-Q, wherein Y is Alanine.
4. The cell culture media according to claim 1 , wherein the only constituent of the set of other L-glutamine sources Qsource is free L-glutamine.
5. The cell culture media according to claim 1 , wherein the total concentration of N-acylated dipeptides Acyl-X-Q is in the range of from 1 mM to 8 mM.
6. The cell culture media according to claim 1 , wherein the molar ratio R=n(Acyl-X-Q)/n(Qsource) is defined to be in the range of 0.04 to 15.67.
7. The cell culture media according to claim 1 , wherein the set of N-acylated dipeptides Acyl-X-Q is defined as follows: One or more of acetyl-A-Q, acetyl-G-Q, acetyl-NQ.
8. The cell culture media according to claim 1 , wherein the set of N-acylated dipeptides Acyl-X-Q is defined as follows: One or more of acetyl-A-Q, acetyl-G-Q.
9. The cell culture media according to claim 1 , wherein the set of N-acylated dipeptides Acyl-X-Q is defined as follows: Acetyl-A-Q.
10. The cell culture media according to claim 1 , wherein:
the cell culture media are chemically defined;
the only constituent of the set of other L-glutamine sources Qsource is dipeptide Y-Q, wherein Y is Alanine;
the molar ratio R=n(Acyl-X-Q)/n(Qsource) is defined to be in the range of 0.04 to 15.67;
the set of N-acylated dipeptides Acyl-X-Q is defined as follows: Acetyl-A-Q; and
the concentration of acetyl-A-Q is in the range of from 1 mM to 8 mM.
11. A process, comprising culturing cells with the cell culture media of claim 1 .
12. A process for manufacturing a cell culture product, the process comprising contacting cells with the cell culture medium of claim 1 .
13. The process according to claim 12 , wherein the cells contacted with the cell culture medium are Chinese hamster ovary (CHO) cells.