期刊
JOURNAL OF BIOTECHNOLOGY
卷 117, 期 3, 页码 243-251出版社
ELSEVIER
DOI: 10.1016/j.jbiotec.2005.01.018
关键词
Arylsulfatase A; Metachromatic Leukodystrophy; purification; enzyme replacement-based therapy
资金
- Telethon [TGM06S01, TGT00B01, TGT06S01] Funding Source: Medline
The production of active Arylsulfatase A is a key step in the development of enzyme replacement therapy for Metachromatic Leukodystrophy. 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 mu g 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. (c) 2005 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据