ALFA-L-IDURONIDASA RECOMBINANT, PROCEDURES TO PRODUCE AND PURIFY THE SAME AND PROCEDURES TO TREAT DISEASES CAUSED BY ITS DEFICIENCY.

摘要

Method for purifying a recombinant aL-iduronidase enzyme or its biologically active fragments or its mutants, to equal or exceed 99% purity, comprising the following steps: (a) recovering and filtering the liquid obtained from a culture of cells transformed with nucleic acids encoding said recombinant aL-iduronidase; (b) adjust the pH of the liquid to an acidic pH, followed by filtration through a filter between 0.2 and 0.54 microns; (c) passing the liquid through a column of blue sepharose FF to capture said a-L-iduronidase; (d) passing the eluate through a column of copper chelating sepharose to remove contaminating proteins; (e) passing the eluate through a phenyl sepharose column to reduce the filtered residual blue Cibacron dye and the copper ions transported from the previous columns; and (f) concentrate and filter the purified recombinant α-L-iduronidase eluted therefrom.