Method for producing a cell capable of high-yield production of heteroproteins
The present invention provides a cell capable of high-yield production of polypeptides and a method for producing the same. The present invention relates to a method for producing a cell capable of high-yield production of a desired polypeptide, wherein a strongly bicarbonate transporter-expressing cell into which DNA encoding the desired polypeptide has been introduced is cultured in the presence of a high concentration of methotrexate and a cell capable of high-yield production of the desired polypeptide is selected from among surviving cells.
1. A method for producing a cell capable of high-yield production of a desired polypeptide, comprising culturing a cell into which a bicarbonate transporter gene has been artificially transferred and into which DNA encoding the desired polypeptide has been introduced, in the presence of a high concentration of methotrexate and selecting a cell capable of high-yield production of the desired polypeptide from among surviving cells.
2. The method according to claim 1 , wherein DNA encoding dihydrofolate reductase is also introduced into the cell into which a bicarbonate transporter gene has been artificially transferred and into which DNA encoding the desired polypeptide has been introduced.
3. The method according to claim 2 , wherein the cell into which a bicarbonate transporter gene has been artificially transferred and into which DNA encoding the desired polypcptide has been introduced is a cell that is co-transformed with one molecule containing DNA encoding the desired polypcptide and DNA encoding dihydrofolate reductase.
4. The method according to claim 3 , wherein the molecule containing DNA encoding the desired polypeptide and DNA encoding dihydrofolate reductase is a vector.
5. The method according claim 1 , wherein the cell into which a bicarbonate transporter gene has been artificially transferred also strongly expresses alanine aminotransferase.
6. A cell produced by a method according to claim 1 .
7. A method for producing a desired polypeptide comprising culturing the cell according to claim 6 .
8. The method according to claim 7 , wherein the desired polypeptide is an antibody.
9. A pharmaceutical product containing a polypeptide produced by the method according to claim 7 .
10. A method for enhancing the amount of polypeptide production by a cell into which a bicarbonate transporter gene has been artificially transferred and into which DNA encoding a desired polypeptide has been introduced, the method comprising treating the cell with a high concentration of methotrexate.
11. The method according to claim 10 . wherein DNA encoding dihydrofolate reductase is also introduced into the cell into which a bicarbonate transporter gene has been artificially transferred and into which DNA encoding the desired polypeptide has been introduced.
12. The method according to claim 11 , wherein the cell into which a bicarbonate transporter gene has been artificially transferred and into which DNA encoding the desired polypeptide has been introduced is a cell that is co-transformed with one molecule containing DNA encoding the desired polypeptide and DNA encoding dihydrofolate reductase.
13. The method according to claim 10 , wherein the strongly cell into which a bicarbonate transporter gene has been artificially transferred also strongly expresses alanine aminotransferase.
14. A method for preparing a cell with a high survival rate in a culture comprising methotrexate at a high concentration, comprising artificially transferring a bicarbonate transporter bene into the cell.
15. A method for producing a cell capable of high-yield production of a desired polypeptide comprising culturing a cell into which a bicarbonate transporter gene has been artificially transferred and into which DNA encoding the desired polypeptide has been introduced in the presence of methotrexate and selecting a cell capable of high-yield production of the desired polypeptide from among surviving cells.
16. A method for enhancing the amount of polypeptide produced by a cell into which a bicarbonate transporter gene has been artificially transferred and into which DNA encoding a desired polypeptide has been introduced, the method comprising treating the cell with methotrexate.