Host cells comprising alpha 1,2 mannosidase and culture methods thereof
Improved host cells and culture methods involving overexpression of MAN1C1 activity to improve protein production are provided.
1. An isolated host cell engineered to overexpress (a) alpha 1,2 mannosidase (MAN1C1) native to the host cell and (b) a glycoprotein of interest, wherein the MAN1C1 polypeptide is expressed at a level that increases specific productivity (pg/cell/day) of said glycoprotein of interest compared to an unaltered host cell.
2. The host cell of claim 1 wherein said host cell has been transfected with a vector comprising a nucleic acid encoding MAN1C1 operably linked to a heterologous expression control sequence.
3. The host cell of claim 1 wherein said host cell comprises a heterologous expression control sequence operably linked to a nucleic acid encoding MAN1C1.
4. The host cell of any of claims 1 - 3 wherein said host cell has been transfected with a vector comprising a nucleic acid encoding said glycoprotein of interest operably linked to a heterologous expression control sequence.
5. The host cell of any of claims 1 - 3 wherein said host cell comprises a heterologous expression control sequence operably linked to a nucleic acid encoding said glycoprotein of interest.
6. The host cell of claim 5 wherein the glycoprotein of interest is an erythropoiesis-stimulating molecule.
7. The host cell of claim 6 wherein the glycoprotein of interest is erythropoietin of SEQ ID NO: 3.
8. The host cell of claim 6 wherein the glycoprotein of interest is darbepoetin of SEQ ID NO: 5.
9. The host cell of claim 5 which is a mammalian cell.
10. The host cell of claim 5 which is a CHO cell.
11. The host cell of claim 5 which is a human cell.
12. The host cell of claim 5 which is a BHK cell.
13. The host cell of claim 5 which is an NS/0 cell.
14. The host cell of claim 5 which is an HT-1080 cell.