Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9604381 | Journal of Biotechnology | 2005 | 9 Pages |
Abstract
To obtain large amounts of purified Arylsulfatase A for therapeutic use, we combined a retroviral expression system with a versatile and rapid purification protocol that can easily and reliably be adapted to high-throughput applications. The purification method consists of an initial ion-exchange DEAE-cellulose chromatography step followed by immuno-affinity purification using a polyclonal antibody against a 29-mer peptide of the Arylsulfatase A sequence. Immuno-adsorbed protein was eluted with a combination of acidic pH and an optimal concentration of the 29-mer peptide. This protocol reproducibly yielded approximately 100 μg of >99% pure human Arylsulfatase A, corresponding to 152 mU of enzyme activity, per liter of culture medium with properties similar to those of human non-recombinant protein.
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Authors
Sabata Martino, Antonella Consiglio, Cristina Cavalieri, Roberto Tiribuzi, Egidia Costanzi, Giovanni Maria Severini, Carla Emiliani, Claudio Bordignon, Aldo Orlacchio,