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US 2008/0293105 A1Published application
Recombinant method for making multimeric proteins
AssigneeICOS Corporation
InventorDaniel S. Allison
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Abstract
The present invention relates to methods for making multimeric proteins comprising fusion of two or more cells expressing a single subunit of the multimeric protein to generate a single hybrid cell expressing the fully assembled multimeric protein.
Claims19
A method for making a multimeric protein comprising the steps of:
- a) transfecting a first host cell with a first plasmid comprising a first polynucleotide encoding a first polypeptide of the multimeric protein, wherein the plasmid is not amplified using an amplifiable marker and wherein the plasmid comprises a selectable marker and a regulatory DNA element which provides increased expression of the first polypeptide;
- b) transfecting a second host cell with a second plasmid comprising a second polynucleotide encoding a second polypeptide of the multimeric protein, wherein the plasmid is not amplified using an amplifiable marker and wherein the plasmid comprises a selectable marker and a regulatory DNA element which provides increased expression of the second polypeptide;
- c) fusing the host cells to make a cell hybrid, wherein the cell hybrid expresses the polypeptides comprising the multimeric protein, and
- d) culturing the cell hybrid in culture media under conditions that permit the expression and association of the polypeptides to form the multimeric protein.
1 more element
A method for making an antibody comprising the steps of:
- a) transfecting a first host cell with a first plasmid comprising a first polynucleotide encoding a heavy chain polypeptide of the antibody, wherein the plasmid is not amplified using an amplifiable marker and wherein the plasmid comprises a selectable marker and a regulatory DNA element which provides increased expression of the heavy chain polypeptide;
- b) transfecting a second host cell with a second plasmid comprising a second polynucleotide encoding a light chain polypeptide of the antibody, wherein the plasmid is not amplified using an amplifiable marker and wherein the plasmid comprises a selectable marker and a regulatory DNA element which provides increased expression of the light chain polypeptide;
- c) fusing the host cells to make a cell hybrid, wherein the cell hybrid expresses the heavy chain polypeptide and the light chain polypeptide, and
- d) culturing the cell hybrid in culture media under conditions that permit the expression and association of the heavy chain and the light chain to form the antibody.
1 more element
Dependencies are inferred from the claim text.