METHOD FOR PRODUCTION OF RECOMBINANT HUMAN ALPHA-GALACTOSIDASE A

摘要

Disclosed is a method for production of recombinant human alpha-galactosidase A (rh alpha-Gal A) in a large scale, with a high purity. The method comprises the steps of (a) culturing rh alpha-Gal A-producing mammalian cells in a serum-free medium, (b) collecting culture supernatant, (c) subjecting the culture supernatant to anion-exchange column chromatography, (d) to hydrophobic column chromatography, (e) to a column chromatography employing as solid phase a material having affinity for phosphate group, (f) to cation-exchange column chromatography, and (g) to gel filtration column chromatography, in the order.

主权项

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 anion-exchange column chromatography or dye affinity column chromatography to collect the recombinant human alpha-Gal A -active fractions, (d) subjecting the fractions collected in step (c) above to hydrophobic column chromatography to collect recombinant human alpha-Gal A-active fractions, (e) subjecting the fractions collected in step (d) above to a column chromatography employing as solid phase a material having affinity for phosphate group to collect the recombinant human alpha-Gal A-active fractions, (f) subjecting the fractions collected in step (e) above to cation-exchange column chromatography to collect the recombinant human alpha-Gal A-active fractions, and (g) subjecting the fractions collected in step (f) above to gel filtration column chromatography to collect the recombinant human alpha-Gal A-active fractions, in the order.