Method of enhancing cell growth using alkyl-amine-n-oxide (AANOx)
View Patent ↗The present invention relates to a method to enhance cell growth in culture comprising adding an alkyl-amine-n-oxide (AANOx), such as dodecyldimethylamine oxide (DDAO), into the culture medium in an amount sufficient to improve cell growth.
1. A method for enhancing expression of recombinant protein in cell culture comprising culturing cells that express the recombinant protein in culture media comprising an amount of dodecyldimethylamine oxide (DDAO) or a related analyte sufficient to improve the growth rate of the cells in culture, wherein the analyte is selected from the group consisting of C10 dimethylamine oxide, C12 dimethyl amine oxide, C14 dimethyl amine oxide, C16 dimethylamine oxide, C18 dimethylamine oxide, dimethyl-tetradecyl-amine-oxide and dimethyl-hexadecyl-amine-oxide.
2. A method for enhancing growth rate of cells in cell culture comprising culturing cells in culture media comprising an amount of dodecyldimethylamine oxide (DDAO) or a related analyte sufficient to improve the growth rate of the cells in culture, wherein the analyte is selected from the group consisting of C10 dimethylamine oxide, C12 dimethyl amine oxide, C14 dimethyl amine oxide, C16 dimethylamine oxide, C18 dimethylamine oxide, dimethyl-tetradecyl-amine-oxide and dimethyl-hexadecyl-amine-oxide.
3. The method of claim 1 wherein the amount of DDAO or a related analyte is between about 4 and about 80 ppb.
4. The method of claim 1 wherein the amount of DDAO or a related analyte is between about 4 and about 50 ppb.
5. The method of claim 1 wherein the amount of DDAO or a related analyte is between about 10 ppb and about 40 ppb.
6. The method of claim 1 wherein DDAO or a related analyte is added over regular time periods to sustain the level of DDAO or a related analyte in the cell media.
7. The method of claim 6 wherein DDAO or a related analyte is added over multiple time periods to sustain the level of DDAO or a related analyte in the cell media.
8. The method of claim 6 wherein DDAO or a related analyte is added continuously over a single time period to sustain the level of DDAO or a related analyte in the cell media.
9. The method of claim 1 wherein the DDAO or a related analyte is not derived from a soy hydrolysate preparation.
10. The method of claim 1 wherein the DDAO or a related analyte is derived from a soy hydrolysate preparation.
11. The method of claim 1 wherein the culture media is animal protein-free media.
12. The method of claim 1 wherein the culture media comprises animal protein.
13. The method of claim 1 wherein the cells are mammalian cells.
14. The method of claim 13 where the cells are selected from the group consisting of BSC cells, LLC-MK cells, CV-1 cells, COS cells, VERO cells, MDBK cells, MDCK cells, CRFK cells, RAF cells, RK cells, TCMK-1 cells, LLCPK cells, PK15 cells, LLC-RK cells, MDOK cells, BHK-21 cells, CHO cells, NS-1 cells, MRC-5 cells, WI-38 cells, BHK cells, 293 cells, and RK cells.
15. The method of claim 13 wherein the cells are CHO cells.
16. The method of claim 1 wherein the recombinant protein is a blood clotting factor.
17. The method of claim 16 where in the blood clotting factor is selected from the group consisting of Factor II, Factor V, Factor VII, Factor VIII, Factor IX, Factor X, Factor XI, von Willebrand Factor, Factor XII and Factor XIII.
18. The method of claim 17 wherein the blood clotting factor is Factor VIII.