Metabolic selection via the serine biosynthesis pathway
AssigneeSigma-Aldrich Co.
InventorsMary Otto, James Ravellette, David Razafsky, Jason A. Gustin, Trissa Borgschulte
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Abstract
The present disclosure provides an isolated mammalian cell comprising a reduced or eliminated expression of Phosphoserine Phosphatase (PSPH). Further provided are methods for preparing such cells and methods for using such cells for the production of recombinant proteins.
Claims22
A method for producing a recombinant protein product, the process comprising
- (a) providing a mammalian cell line engineered to have reduced or eliminated expression of endogenous phosphoserine phosphatase (PSPH);
- (b) introducing into the mammalian cell line a polynucleotide, wherein the polynucleotide encodes a functional PSPH gene and the recombinant protein;
- (c) culturing the cell line;
- and
- (d) purifying the recombinant protein to form the recombinant protein product.
A genetically engineered mammalian cell line for use in a biologic production system, wherein the mammalian cell line is engineered to have reduced or eliminated expression of endogenous PSPH.
A polynucleotide comprising a nucleic acid sequence encoding a functional PSPH and at least one recombinant protein of interest.
A polynucleotide comprising
- a) a nucleic acid sequence encoding a functional PSPH;
- b) a nucleic acid sequence encoding a functional GS and/or ASNS;
- and
- c) a nucleic acid sequence encoding a mutation in the ASNS and/or PSPH coding sequence that attenuates the activity of either one or both of the enzymes
- d) a nucleic acid sequence encoding a recombinant protein of interest.
Dependencies are inferred from the claim text.