Patent ID: 9279109
Filing Date: 2016-03-08
CPC Classification: A61P,C12N,C12Y

Claim Text:
1. A method for producing recombinant human wild-type iduronate 2-sulfatase (rhI2S) consisting of the 26 th to 550 th amino acids of the amino acid sequence set forth in SEQ ID NO:10, wherein the method comprises: (a) culturing rhI2S-producing mammalian cells in a serum-free medium so as to allow the cells to secrete the rhI2S in the medium, wherein the cells are those that have been transformed to express the gene encoding the rhI2S by introduction of the gene using an expression vector in which the gene is incorporated, (b) collecting culture supernatant by removing the cells from the culture that is obtained in step (a) above, (c) subjecting the culture supernatant collected in step (b) above to cation-exchange column chromatography to collect rhI2S-active fractions, (d) subjecting the fractions collected in step (c) above to dye affinity column chromatography to collect rhI2S-active fractions, (e) subjecting the fractions collected in step (d) above to anion-exchange column chromatography to collect rhI2S-active fractions, (f) subjecting the fractions collected in step (e) above to a column chromatography employing as a solid phase, a material having affinity for phosphate group to collect rhI2S-active fractions, and (g) subjecting the fractions collected in step (1) above to gel filtration column chromatography to collect rhI2S-active fractions, in the respective order.