Modified mammalian cells
AssigneeGenentech, Inc.
InventorsMichael Wilson LAIRD, Shahram MISAGHI, Amy SHEN, Anthony TOMLINSON, Inn Huam YUK
Life of this patent
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Abstract
The present disclosure relates to methods, cells, and compositions for producing a product of interest, e.g., a recombinant protein. In particular, the present disclosure provides improved mammalian cells expressing the product of interest, where the cells (e.g., Chinese Hamster Ovary (CHO) cells) have reduced or eliminated activity, e.g., expression, of certain host cell proteins, e.g., enzymes including, but not limited to, certain lipases, esterases, and/or hydrolases.
Claims27
A recombinant host cell, wherein the cell is modified to reduce or eliminate the activity of one or more enzyme relative to the activity of the enzyme in an unmodified cell.
-16. (canceled)
A method for reducing enzymatic activity in a cell, comprising:
- (a) knocking down or knocking out the expression of LPL;
- PLBL2/PLBD2;
- LIPA;
- PLAA;
- PLD3;
- LPLA2;
- PLCB1;
- PLCD1;
- DDHD1;
- LYPLAL1;
- PLA2G12A;
- PRDX6;
- SMPD1;
- PPT1;
- IAH1;
- OTUB1;
- LYPLA2;
- ACOT13;
- FASN;
- PLA2G7;
- USP5;
- ASAH1;
- LMF1;
- APOC2;
- HACH;
- CES1F/CESB1L;
- LYPLA1;
- CES1;
- PLA1A;
- and/or SIAE;
- (b) modulating a cell culture process and/or media composition, wherein modulating a cell culture process and/or media composition results in reduced activity of LPL PLBL2/PLBD2 LIPA PLAA PLD3 LPLA2 PLCB1;
- PLCD1;
- DDHD1;
- LYPLAL1;
- PLA2G12A;
- PRDX6;
- SMPD1;
- PPT1;
- IAH1;
- OTUB1;
- LYPLA2;
- ACOT13;
- FASN;
- PLA2G7;
- USP5;
- ASAH1;
- LMF1;
- APOC2;
- HACH;
- CES1F/CES-B1L;
- LYPLA1;
- CES1;
- PLA1A;
- and/or SIAE;
- (c) selecting cells with reduced activity of LPL;
- PLBL2/PLBD2;
- LIPA;
- PLAA;
- PLD3;
- LPLA2;
- PLCB1;
- PLCD1;
- DDHD1;
- LYPLAL1;
- PLA2G12A;
- PRDX6;
- SMPD1;
- PPT1;
- IAH1;
- OTUB1;
- LYPLA2;
- ACOT13;
- FASN;
- PLA2G7;
- USP5;
- ASAH1;
- LMF1;
- APOC2;
- HACH;
- CES1F/CES-B1L;
- LYPLA1;
- CES1;
- PLA1A;
- and/or SIAE or
- (d) altering the gene encoding one or more of LPL;
- PLBL2/PLBD2;
- LIPA;
- PLAA;
- PLD3;
- LPLA2;
- PLCB1;
- PLCD1;
- DDHD1;
- LYPLAL1;
- PLA2G12A;
- PRDX6;
- SMPD1;
- PPT1;
- IAH1;
- OTUB1;
- LYPLA2;
- ACOT13;
- FASN;
- PLA2G7;
- USP5;
- ASAH1;
- LMF1;
- APOC2;
- HACH;
- CES1F/CES-B1L;
- LYPLA1;
- CES1;
- PLA1A;
- and/or SIAE.
-25. (canceled)
A method for reducing or eliminating enzymatic activity in a cell, comprising administering to the cell a genetic engineering system, wherein the genetic engineering system alters the nucleic acid sequence encoding one or more of LPL;
- PLBL2/PLBD2;
- LIPA;
- PLAA;
- PLD3;
- LPLA2;
- PLCB1;
- PLCD1;
- DDHD1;
- LYPLAL1;
- PLA2G12A;
- PRDX6;
- SMPD1;
- PPT1;
- IAH1;
- OTUB1;
- LYPLA2;
- ACOT13;
- FASN;
- PLA2G7;
- USP5;
- and ASAH1 so that the one or more of LPL;
- PLBL2/PLBD2;
- LIPA;
- PLAA;
- PLD3;
- LPLA2;
- PLCB1;
- PLCD1;
- DDHD1;
- LYPLAL1;
- PLA2G12A;
- PRDX6;
- SPD1;
- PPT1;
- IAH1;
- OTUB1;
- LYPLA2;
- ACOT13;
- FASN;
- PLA2G7;
- USP5;
- ASAH1;
- LMF1;
- APOC2;
- HACH;
- CES1F/CES-B1L;
- LYPLA1;
- CES1;
- PLA1A;
- and/or SIAE have reduced or eliminated enzymatic activity.
-44. (canceled)
A method of producing a product of interest comprising culturing mammalian cells expressing the product of interest, wherein the mammalian cells express the product of interest and have reduced or eliminated activity of one or more of the following LPL;
- PLBL2/PLBD2;
- LIPA;
- PLAA;
- PLD3;
- LPLA2;
- PLCB1;
- PLCD1;
- DDHD1;
- LYPLAL1;
- PLA2G12A;
- PRDX6;
- SMPD1;
- PPT1;
- IAH1;
- OTUB1;
- LYPLA2;
- ACOT13;
- FASN;
- PLA2G7;
- USP5;
- ASAH1;
- LMF1;
- APOC2;
- HACH;
- CES1F/CES-B1L;
- LYPLA1;
- CES1;
- PLA1A;
- and/or SIAE.
-59. (canceled)
-117. (canceled)
Dependencies are inferred from the claim text.