Methods of protein production using anti-senescence compounds
AssigneeWyeth
InventorsJose Manuel Gomes, Yen-Tung Luan, Gregory Walter Hiller, Wenge Wang
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Abstract
Methods of producing a protein in cell culture comprising an anti-senescence compound, such as the antioxidant carnosine, are provided. According to teachings of the present invention, cells grown in a cell culture medium comprising an anti-senescence compound exhibit increased viability and productivity. Furthermore, cell cultures grown in the presence of an anti-senescence compound exhibit decreased levels of high molecular weight aggregates in the cell culture medium.
Claims30
A method of producing a TNFR fusion protein in cell culture comprising steps of:
- culturing mammalian cells that contain a gene encoding the TNFR fusion protein, which gene is expressed under conditions of cell culture, in a cell culture medium comprising an anti-senescence compound, wherein the anti-senescence compound is selected from the group consisting of carnosine, acetyl-carnosine, homo-carnosine, anserine, and beta-alanine and combinations thereof;
- and
- maintaining the culture under conditions and for a time sufficient to permit expression of the protein;
- and
- harvesting the cell culture medium, which contains the protein;
- and wherein the mammalian cells are Chinese hamster ovary (CHO) cells.
A method for producing a TNFR fusion protein comprising steps of:
- culturing mammalian cells that contain a gene encoding the TNFR fusion protein in a cell culture medium, which gene is expressed under conditions of cell culture, at a first temperature or temperature range conducive for cell growth during a growth phase;
- shifting the temperature or temperature range of the cell culture medium to a second temperature or temperature range conducive for protein production;
- culturing the host cells in the cell culture medium at the second temperature or temperature range through a transition phase and into a production phase;
- wherein an anti-senescence compound is added to the cell culture, wherein the anti-senescence compound is selected from the group consisting of carnosine, acetyl-carnosine, homo-carnosine, anserine, and beta-alanine and combinations thereof;
- and
- harvesting the cell culture medium, which contains the protein;
- wherein the mammalian cells are Chinese hamster ovary (CHO) cells.
A method of decreasing accumulation of high molecular weight aggregates during production of a TNFR fusion protein in cell culture comprising steps of:
- culturing Chinese hamster ovary (CHO) cells that contain a gene encoding the TNFR fusion protein, which gene is expressed under conditions of cell culture, in a cell culture medium comprising;
- and
- maintaining the culture under conditions and for a time sufficient to permit expression of the protein;
- and
- harvesting the cell culture medium, which contains the protein.
A method of decreasing accumulation of acidic species during production of a TNFR fusion protein in cell culture comprising steps of:
- culturing Chinese hamster ovary (CHO) cells that contain a gene encoding the TNFR fusion protein, which gene is expressed under conditions of cell culture, in a cell culture medium comprising carnosine;
- and
- maintaining the culture under conditions and for a time sufficient to permit expression of the protein.
Dependencies are inferred from the claim text.