Fibroblast growth factor muteins with increased activity
View Patent ↗The present invention provides FGF-2 polypeptides that differ from the wild-type FGF-2 (SEQ ID NO:1) at least at amino acid position 56, 102 and 119, wherein the differences are Q56I, N102G, K119N substitutions, leading to FGF-2 polypeptides with higher thermostability, higher biological activity and higher resistance to proteolytic degradation. These polypeptides can be used in cell culture media.
1. A fibroblast growth factor-two (FGF-2) polypeptide that differs from the wild-type FGF-2 (SEQ ID NO:1) at least at amino acid positions 56, 102 and 119, wherein the differences are Q56I, N102G and K119N substitutions, and wherein said FGF-2 polypeptide has a higher thermostability, higher biological activity and higher resistance to proteolytic degradation than wild-type FGF-2.
2. An FGF-2 polypeptide according to claim 1 , wherein said FGF-2 polypeptide has the sequence of SEQ ID NO:4.
3. An FGF-2 polypeptide according to claim 1 , wherein said FGF-2 polypeptide additionally differs from the wild-type FGF-2 at positions 69 and 87, wherein the additional differences are C69S and C87S substitutions.
4. An FGF-2 polypeptide according to claim 3 , wherein said FGF-2 polypeptide has the sequence of SEQ ID NO:6.
5. A method for culturing FGF-2 sensitive cells in a medium, the method comprising the step of cultivation of FGF-2 sensitive cells, wherein said medium comprises an FGF-2 polypeptide according to claim 1 .
6. A method according to claim 5 , wherein said sensitive cells are selected from the group consisting of pluripotent cells, embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells, bone marrow stromal cells and neural stem cells.