On-Column Refolding and Purification of Recombinant Human Interleukin-1 Receptor Antagonist (rHuIL-1ra) Expressed as Inclusion Body in Escherichia coli

Authors: Tan, Haidong¹; Dan, Guoping²; Gong, Huiying²; Cao, Lijun²

Source: Biotechnology Letters, Volume 27, Number 16, August 2005 , pp. 1177-1182(6)

Publisher: Springer

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Abstract:

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.

Keywords: human interleukin-1 receptor antagonist; inclusion body; on-column refolding

Document Type: Research article

DOI: http://dx.doi.org/10.1007/s10529-005-8655-5

Affiliations: 1: Dalian Institute of Chemical Physics, CAS, 457 Zhongshan Road, 116023, Dalian, P.R. China, Email: hdt@dicp.ac.cn 2: Yushui Biopharmaceutical Co., Ltd, 200400, Chongqing, P.R. China,

Publication date: 2005-08-01