Proteins with modified glycosylation and methods of production thereof
View Patent ↗In one aspect, the disclosure provides proteins with modified glycosylation and methods of their production. In aspect, the disclosure provides transgenic animals and cells for the production of proteins with modified glycosylation. In some embodiment, the modified glycosylation is increased sialylation.
1. A method of producing a therapeutic protein with increased sialylation, the method comprising:
providing a transgenic non-human mammal that has been modified to transgenically express a sialyl transferase and a therapeutic protein in mammary gland epithelial cells of the transgenic non-human mammal,
wherein a gene encoding the sialyl transferase and a gene encoding the therapeutic protein are each integrated into the genome of the transgenic non-human mammal, and wherein the expression of the sialyl transferase and the therapeutic protein is under the control of at least one milk promoter;
expressing the sialyltransferase and the therapeutic protein; and
harvesting the therapeutic protein with increased sialylation from the mammary gland of the transgenic non-human mammal;
wherein the sialylation of the therapeutic protein is increased by at least 10% compared to a therapeutic protein produced in a transgenic non-human mammal that has not been modified to transgenically express a sialyl transferase.
2. The method of claim 1 , wherein the therapeutic protein with increased sialylation is a human protein.
3. The method of claim 1 , wherein the protein with increased sialylation is an antibody.
4. The method of claim 1 , wherein the protein with increased sialylation is antithrombin.
5. The method of claim 1 , wherein the protein with increased sialylation is alpha-1 antitrypsin.
6. A transgenic non-human mammal that transgenically expresses a sialyl transferase and a therapeutic protein in mammary gland epithelial cells of the transgenic non-human mammal,
wherein a gene encoding the sialyl transferase and a gene encoding the therapeutic protein are each integrated into the genome of the transgenic non-human mammal,
wherein expression of the sialyl transferase and the therapeutic protein is under the control of at least one milk promoter, and
wherein the sialylation of the therapeutic protein produced in the transgenic non-human mammal is increased at least 10% compared to a therapeutic protein produced in a transgenic non-human mammal that has not been modified to transgenically express the sialyl tranferase.
7. The transgenic non-human mammal of claim 6 , wherein the therapeutic protein is a human protein.
8. The transgenic non-human mammal of claim 6 , wherein the therapeutic protein is an antibody.
9. The transgenic non-human mammal of claim 6 , wherein the therapeutic protein is antithrombin.
10. The transgenic non-human mammal of claim 6 , wherein the therapeutic protein is alpha-1 antitrypsin.
11. The method of claim 1 , wherein the transgenic mammal is an ungulate.
12. The method of claim 11 , wherein the ungulate is a goat.
13. The method of claim 1 , wherein the milk promoter is a goat beta casein promoter.
14. The method of claim 1 , wherein the sialyl transferase is beta-galactosamide alpha-2,6-sialyltranferase and/or beta-galactosamide alpha-2,3-sialyltranferase.