On-column refolding and purification of recombinant human interleukin-1 receptor antagonist (rHuIL-1ra) expressed as inclusion body in Escherichia coli

被引：7
|
作者：
Tan, HD
Dan, GP
Gong, HY
Cao, LJ
机构：
[1] Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
[2] Yushui Biopharmaceut Co Ltd, Chongqing 200400, Peoples R China
来源：
BIOTECHNOLOGY LETTERS | 2005年 / 27卷 / 16期
关键词：
human interleukin-1 receptor antagonist; inclusion body; on-column refolding;
D O I：
10.1007/s10529-005-8655-5
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Recombinant human interleukin-1 receptor antagonist (rHuIL-1ra) was produced in E. coli as an inclusion body. rHuIL-1ra was purified to Over 98% purity by anion exchange chromatography after on-column refolding. The optimized processes produced more than 2 g pure refolded rHuIL-1ra per 1 l culture, corresponding to a 44% recovery, without an intermediate dialysis step. Refolded rHuIL-1ra had full biological activity with the MTT assay. An intramolecular disulfide linkage in the oxidized recombinant protein was suggested by data from HPLC and non-reducing SDS-PAGE.
引用
收藏
页码：1177 / 1182
页数：6
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