Patent ID: 9617529
Date: 2017-04-11
CPC Classifications: C12N

Claim:
1. A method for production of a recombinant human alpha-Gal A comprising the steps of: (a) culturing recombinant human alpha-Gal A-producing mammalian cells in a serum-free medium to let them secrete the recombinant human alpha-Gal A in the medium, (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 an anion-exchange column chromatography to collect first recombinant human alpha-Gal A-active fractions, (d) subjecting the first fractions collected in step (c) above to a hydrophobic column chromatography to collect second recombinant human alpha-Gal A-active fractions, wherein the hydrophobic column chromatography employs a hydrophobic ligand, wherein the hydrophobic ligand contains a phenyl group, (e) subjecting the second fractions collected in step (d) above to a column chromatography employing as solid phase a material having affinity for phosphate group to collect third recombinant human alpha-Gal A-active fractions, (f) subjecting the third fractions collected in step (e) above to a cation-exchange column chromatography to collect fourth recombinant human alpha-Gal A-active fractions, and (g) subjecting the fourth fractions collected in step (f) above to a gel filtration column chromatography to collect fifth recombinant human alpha-Gal A-active fractions, in the order listed.