Cell culture media containing combinations of proteins
The present invention relates to cell culture media supplements or complete media compositions comprising plant-produced heterologous recombinant human albumin, as well as methods of making the cell culture media, and methods of using the supplemented cell culture media to improve viability, productivity, and growth characteristics of cultured cells.
1. A cell culture media supplement or complete media composition for improving growth, viability, and/or productivity of cultured cells as compared to cells grown without said supplement or complete media composition, said cell culture media supplement or complete media composition comprising a plant-produced heterologous protein, wherein said plant-produced heterologous protein is recombinant human albumin, said recombinant human albumin comprising:
i) less than 1 EU of endotoxin/mg of albumin; and
ii) less than 10 ppm detergent.
2. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin is a sterile liquid or powder.
3. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin has a molecular weight of 65-75 kDa.
4. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin has an isoelectric focusing point of 5.3.
5. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin comprises albumin and fragments thereof provided that the fragments retain the biological or therapeutically beneficial activity of said albumin.
6. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin further comprises albumin dimers having a molecular weight of about 130 kDa as detected by SDS-PAGE.
7. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin comprises less than 2% aggregates at around 250 KDa as detected by SDS-PAGE.
8. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin has an amino acid sequence that is homologous to native human albumin.
9. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin is recombinant human serum albumin.
10. The cell culture media supplement or complete media composition of claim 9 , wherein said recombinant human serum albumin has an amino acid sequence that is at least 80% identical to secreted human serum albumin comprising the amino acid sequence of SEQ ID NO: 1.
11. The cell culture media supplement or complete media composition of claim 9 , wherein said recombinant human serum albumin comprises the amino acid sequence of SEQ ID NO: 12.
12. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin comprises one or more bound lipids from a plant.
13. The cell culture media supplement or complete media composition of claim 1 , wherein said recombinant human albumin is a protein extract of a transgenic monocot plant.
14. The cell culture media supplement or complete media composition of claim 13 , wherein said recombinant human albumin comprises a monocot seed component.
15. The cell culture media supplement or complete media composition of claim 14 , wherein said monocot seed component is a plant carbohydrate or plant lipid.
16. The cell culture media supplement or complete media composition of claim 13 , wherein said recombinant human albumin is a protein extract of a transgenic seed.
17. The cell culture media supplement or complete media composition of claim 13 , wherein said recombinant human albumin is a protein extract of a transgenic rice seed.
18. The cell culture media supplement or complete media composition of claim 17 , wherein said rice is Oryza sativa.
19. The cell culture media supplement or complete media composition of claim 1 , wherein said supplement or complete media composition is free of serum derived albumin and yeast derived albumin.
20. The cell culture media supplement or complete media composition of claim 1 , further comprising at least one additional protein.
21. The cell culture media supplement or complete media composition of claim 20 , wherein said additional protein is selected from the group consisting of growth factors, lactoferrin, transferrin, insulin, growth hormone, fibronectin attachment factor, lamin attachment factor, collagenase, platelet derived growth factor, brain-derived neurotrophic factor, glial-derived neurotrophic factor, thymic factors, haptocorin, lactahedrin, lactoperoxidase, alpha-fetoprotein, immunoglobin, and alpha-lactalbumin.
22. The cell culture media supplement or complete media composition of claim 20 , wherein said additional protein is a heterologous plant-produced protein.
23. The cell culture media supplement or complete media composition of claim 20 , further comprising selenium and ethanolamine.
24. A cell culture media supplement or complete media composition for improving growth, viability, and/or productivity of cultured cells as compared to cells grown without said supplement or complete media composition, said cell culture media supplement or complete media composition comprising a plant-produced heterologous protein, wherein said plant-produced heterologous protein is recombinant human albumin, said recombinant human albumin comprising:
i) less than 1 EU of endotoxin/mg of albumin; and
ii) a monocot seed component, wherein said monocot seed component is a plant carbohydrate or plant lipid.
25. The cell culture media supplement or complete media composition of claim 24 , wherein said a recombinant human albumin has a purity of greater than 95%.
26. A cell culture media supplement or complete media composition for improving the growth of a cell in cell culture comprising:
a recombinant mammalian albumin wherein said albumin is:
i) produced in a transgenic plant; and
ii) has less than 1 EU of endotoxin/mg of albumin.
27. The composition of claim 26 , wherein said albumin is recombinant human serum albumin.
28. The composition of claim 26 , wherein said transgenic plant is a transgenic grain.
29. The composition of claim 28 , wherein the transgenic grain is transgenic rice.