Cells for producing recombinant iduronate-2-sulfatase
AssigneeShire Human Genetic Therapies, Inc.
InventorsFerenc Boldog, Michael Heartlein
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Abstract
The present invention provides, among other things, methods and compositions for production of recombinant I2S protein with improved potency and activity using cells co-express I2S and FGE protein. In some embodiments, cells according to the present invention are engineered to simultaneously over-express recombinant I2S and FGE proteins. Cells according to the invention are adaptable to various cell culture conditions. In some embodiments, cells of the present invention adaptable to a large-scale suspension serum-free culture.
Claims30
A cell comprising
- a first nucleic acid encoding an iduronate-2-sulfatase (I2S) protein comprising an amino acid sequence at least 70% identical to SEQ ID NO:1;
- and
- a second nucleic acid encoding a formylglycine generating enzyme (FGE) protein comprising an amino acid sequence at least 70% identical to SEQ ID NO:5,
- wherein the first and/or the second nucleic acid are exogenous and wherein the cell, once cultivated under a cell culture condition, produces I2S protein comprising at least about 70% conversion of the cysteine residue corresponding to Cys59 of SEQ ID NO:1 to Cα-formylglycine (FGly).
A cell comprising
- a first nucleic acid encoding an iduronate-2-sulfatase (I2S) protein comprising an amino acid sequence at least 70% identical to SEQ ID NO:1;
- and
- a second nucleic acid encoding a formylglycine generating enzyme (FGE) protein comprising an amino acid sequence at least 70% identical to SEQ ID NO:5,
- wherein the first and/or the second nucleic acid are exogenous and wherein the cell, once cultivated under a cell culture condition, produces I2S protein comprising at least about 60% conversion of the cysteine residue corresponding to Cys59 of SEQ ID NO:1 to Cα-formylglycine (FGly) and at a specific productivity rate of great than about 30 picogram/cell/day.
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A method of producing recombinant iduronate-2-sulfatase (I2S) protein comprising cultivating a cell of any one of the preceding claims under conditions such that the recombinant I2S and FGE proteins are co-expressed in the cell.
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Dependencies are inferred from the claim text.