title: On-column refolding and purification of recombinant human interleukin-1 receptor antagonist (rHuIL-1ra) expressed as inclusion body in Escherichia coli.
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.
authors.affiliation: Chongqing Duotai Company, Chongqing, P.R. China. shugangli@163.com [corrected]
authors.forename: Haidong
authors.initials: H
authors.lastname: Tan
authors.forename: Guoping
authors.initials: G
authors.lastname: Dan
authors.forename: Huiying
authors.initials: H
authors.lastname: Gong
authors.forename: Lijun
authors.initials: L
authors.lastname: Cao
mesh_terms: D002852:Chromatography, Ion Exchange / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry / Q000378:metabolism*; D053590:Interleukin 1 Receptor Antagonist Protein; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012795:Sialoglycoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D011994:Recombinant Proteins; D012795:Sialoglycoproteins