Patent ID: 8536315

Claim:
A process for production of a purified recombinant mature polypeptide with arylsulfatase A (ASA) activity in a continuous cell culture system, the process comprising: i) continuously culturing a mammalian cell capable of secreting said mature polypeptide into a liquid culture medium; and ii) purifying the mature polypeptide by a purification process comprising one or more steps of affinity chromatography and/or ion exchange chromatography, wherein the process of (ii) comprises a polishing step including a passive step, wherein the mature polypeptide with arylsulfatase A activity passes through a cation exchange chromatography resin or membrane and/or affinity chromatography resin without binding thereto and an active step, wherein the mature polypeptide with arylsulfatase A activity is detained within and subsequently eluted from an anion exchange membrane or resin, and wherein the cation exchange chromatography resin or membrane and the anion exchange resin or membrane are coupled or connected in a series, wherein said mammalian cell comprises a nucleotide sequence which encodes a polypeptide comprising (a) SEQ ID NOs:2 or 4 or (b) a mutant sequence at least 95% identical to SEQ ID NOs:2 or 4, and wherein said mature polypeptide is a post-translationally modified product thereof that is produced and secreted by said cell, and has arylsulfatase A activity, and wherein the continuous culturing is for a period of at least one week.