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AssigneeMerck Sharp & Doheme Corp.
InventorsBo Jiang, Stephanie A. Nelson, Rebecca D. Argyros, Dongxing Zha
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Lower eukaryote host cells in which expression of the endogenous protein mannosyltransferase 2 (PMT2) gene has been disrupted by introducing a nucleic acid molecule encoding a Pmt2p protein having a mutation in a conserved region of the protein. The mutation confers to the host cell resistance to PMT inhibitors, which are used to reduce the amount of O-glycosylation of recombinant proteins produced by the host cells but which also have the effect of reducing the robustness of the host cells during fermentation. When host cells that express the mutated PMT2 gene but not the endogenous Pmt2p are cultivated in the presence of a P MT inhibitor, the host cells display an increase in cellular robustness during fed-batch fermentation and express recombinant pro teins in high yield while the amounts O-glycosylation are similar to that produced under similar conditions by host cells that express only the endogenous P MT2 gene.
A lower eukaryote host cell comprising a disruption in the expression of the endogenous protein mannosyltransferases 2 (PMT2) gene and a nucleic acid molecule encoding a mutant Pmt2p comprising at least one amino acid substitution, deletion, or insertion in the region of the Pmt2p protein comprising a conserved region having at least 80%, 90%, or 95% identity to the amino acid sequence of SEQ ID NO:9.
A method for producing a recombinant heterologous protein in a lower eukaryote comprising:
A method for producing a recombinant heterologous protein in a lower eukaryote comprising:
A process for producing recombinant therapeutic proteins comprising:
Dependencies are inferred from the claim text.