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Last Updated: April 26, 2024

Claims for Patent: 7,091,004


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Summary for Patent: 7,091,004
Title:Regulation of translation for recombinant protein production
Abstract: The invention provides cells that produce increased levels of recombinant protein by modulating the activity of translational regulator gene products that are downstream targets of PKB alpha, methods of making such cells, and methods of using such cells. Such translational regulator gene products include 4E-BP1 and mTOR.
Inventor(s): Morris; Arvia E. (Seattle, WA), Wong-Madden; Sharon T. (Bellevue, WA)
Assignee: Immunex Corporation (Seattle, WA)
Application Number:10/165,591
Patent Claims:1. An isolated eukaryotic host cell genetically engineered to express a protein of interest wherein the protein of interest is expressed as an extracellular product and wherein the eukaryotic cell has been genetically engineered to also express mammalian Target of Rapamycin (mTOR).

2. The eukaryotic host cell of claim 1 wherein the host cell is further genetically engineered to express a first selectable marker.

3. The eukaryotic host cell of claim 1 wherein the host cell is a mammalian cell.

4. The eukaryotic host cell of claim 3 wherein the host cell is selected from the group consisting of CHO, VERO, BHK, HeLa, CV1, COS, MDCK, 293, 3T3, myeloma, PC12 and WI38 cells.

5. The eukaryotic host cell of claim 1 wherein the host cell is adapted to grow in serum-free medium.

6. The eukaryotic host cell of claim 5 wherein the host cell is adapted to grow in protein-free medium.

7. The eukaryotic host cell of claim 1 wherein the protein of interest is selected from the group consisting of a soluble TNF receptor, a soluble IL-4 receptor, a soluble IL-1 type II receptor, a soluble Flt3 ligand, a soluble CD40 ligand, CD39, CD30, CD27, a TEK/Ork, IL-15, a soluble IL-15 receptor, Ox 40, GM-CSF, RANKL, RANK, TRAIL, a soluble TRAIL receptor, tissue plasminogen activator, Factor VIII, Factor IX, apolipoprotein E, apolipoprotein A-I, an IL-2 receptor, an IL-2 antagonist, alpha-1 antitrypsin, calcitonin, growth hormone, insulin, insulinotropin, insulin-like growth factors, parathyroid hormone, interferons, superoxide dismutase, glucagon, an erythropoeitin, an antibody, glucocerebrosidase, an Fc-fusion protein, globins, nerve growth factors, interleukins, and colony stimulating factors.

8. A method of producing a protein of interest, the method comprising culturing an isolated eukaryotic host cell genetically engineered to express a protein of interest wherein the protein of interest is an extracellular product and wherein the host cell is genetically engineered to also express mTOR, under conditions such that the protein of interest is expressed.

9. The method of claim 8 further comprising collecting the protein of interest.

10. The method of claim 8 wherein the eukaryotic host cell is transiently transfected.

11. The method of claim 8 wherein the eukaryotic host cell is stably transformed.

12. The method of claim 8 wherein the eukaryotic host cell is selected from the group consisting of CHO, VERO, BHK, HeLa, CV1, COS, MDCK, 293, 3T3, myeloma, PC12 and WI38 cells.

13. The method of claim 8 wherein the protein of interest is selected from the group consisting of a soluble TNF receptor, a soluble IL-4 receptor, a soluble IL-1 type II receptor, a soluble Flt3 ligand, a soluble CD40 ligand, CD39, CD30, CD27, a TEK/Ork, IL-15, a soluble IL-15 receptor, Ox 40, GM-CSF, RANKL, RANK, TRAIL, a soluble TRAIL receptor, tissue plasminogen activator, Factor VIII, Factor IX, apolipoprotein E, apolipoprotein A-I, an IL-2 receptor, an IL-2 antagonist, alpha-1 antitrypsin, calcitonin, growth hormone, insulin, insulinotropin, insulin-like growth factors, parathyroid hormone, interferons, superoxide dismutase, glucagon, an erythropoeitin, an antibody, glucocerebrosidase, an Fc-fusion protein, globins, nerve growth factors, interleukins, and colony stimulating factors.

14. The method of claim 8 wherein the eukaryotic host cell is adapted to grow in serum-free medium.

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