Modified cells for the production of a recombinant product of interest
AssigneeGenentech, Inc.
InventorsShahram MISAGHI, Brian Megia Castellano, Danming Tang, Avi J. Ashkenazi, Bradley R. Snedecor
Life of this patent
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Abstract
The present disclosure relates to cells (e.g., Chinese Hamster Ovary (CHO) cells) that are modified to reduce or eliminate the expression of certain endogenous cellular proteins, and methods of using such cells in the production of a recombinant product of interest, e.g., a recombinant protein, a recombinant viral particle, or a recombinant viral vector. These modifications were specifically chosen to generate engineered cells with desired traits in several key areas, including improved cell culture performance (e.g., higher viability and product titers).
Claims36
A modified cell, wherein the cell is modified to reduce or eliminate the expression of two or more endogenous proteins relative to the expression of the endogenous proteins in an unmodified cell, wherein:
- (a) one or more of the endogenous proteins having reduced or eliminated expression promotes apoptosis of the modified cell during cell culture;
- and
- (b) one or more of the endogenous proteins having reduced or eliminated expression regulates the unfolded protein response (UPR).
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A composition comprising a modified cell, wherein the cell is modified to reduce or eliminate the expression of two or more endogenous proteins relative to the expression of the endogenous proteins in an unmodified cell, wherein:
- (a) one or more of the endogenous proteins having reduced or eliminated expression promotes apoptosis of the modified cell during cell culture;
- and
- (b) one or more of the endogenous proteins having reduced or eliminated expression regulates the unfolded protein response (UPR).
A method of producing a recombinant product of interest comprising:
- (a) culturing a modified cell, wherein the cell is modified to reduce or eliminate the expression of two or more endogenous proteins relative to the expression of the endogenous proteins in an unmodified cell, wherein:
- (i) one or more of the endogenous proteins having reduced or eliminated expression promotes apoptosis of the modified cell during cell culture;
- and
- (ii) one or more of the endogenous proteins having reduced or eliminated expression regulates the unfolded protein response (UPR);
- and
- (b) recovering the recombinant product of interest from a cultivation medium or the modified cells, wherein the modified cells expressing the recombinant product of interest exhibit reduced or eliminated expression of BAX, BAK, and PERK.
A method for producing a modified cell, comprising:
- (a) applying a nuclease-assisted and/or nucleic acid targeting BAX, BAK, and PERK, in the cell to reduce or eliminate the expression of said endogenous genes, and
- (b) selecting the modified cell wherein the expression of said endogenous genes have been reduced or eliminated as compared to an unmodified cell.
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Dependencies are inferred from the claim text.