Production cell line enhancers
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsGang CHEN, Darya BURAKOV, Dipali DESHPANDE
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Abstract
The present invention relates to discovery of the ectopic expression of EDEM2 in a production cell to improve the yield of a useful multi-subunit protein. Thus, the present invention provides for production cell lines, such as the canonical mammalian biopharmaceutical production cell, the CHO cell, containing recombinant polynucleotides encoding EDEM2. Also disclosed is a production cell containing both an EDEM2-encoding polynucleotide as well an XBP1-encoding polynucleotide. Improved titers of antibodies produced by these cell lines are disclosed, as well as the improved cell densities attained by these cells in culture.
Claims36
A cell comprising:
- a. a first polynucleotide comprising a nucleic acid sequence encoding a first production enhancer protein comprising an amino acid sequence that is at least 86% identical to SEQ ID NO:9;
- and
- b. a second polynucleotide comprising a nucleic acid encoding a multi-subunit protein comprising an amino acid sequence of SEQ ID NO:44.
A method of making a multi-subunit protein comprising:
- a. culturing in media a plurality of cells descended by clonal expansion from a cell comprising:
- i. a first polynucleotide comprising a nucleic acid encoding a first production enhancer protein comprising an amino acid sequence that is at least 86% identical to SEQ ID NO:9, and
- ii. a second polynucleotide comprising a nucleic acid encoding a multi-subunit protein comprising an amino acid sequence of SEQ ID NO:44;
- b. allowing the cells to secrete the multi-subunit protein into the media at a rate greater than 9 pg/cell/day;
- and
- c. purifying the multi-subunit protein from the media by affinity chromatography, ion exchange chromatography, or a combination of affinity chromatography and ion exchange chromatography.
Dependencies are inferred from the claim text.