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Evaluating the autoinduction expression system and one‐step purification for high‐level expression and purification of gallbladder‐derived rhIL‐1Ra
Author(s) -
Bashir Hamid,
Ahmed Nadeem,
Khan Mohsin Ahmad,
Zafar Ahmad Usman,
Tahir Saad,
Kanwal Hina,
Khan Faidad,
Rahman Zia ur,
Husnain Tayyab
Publication year - 2016
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1463
Subject(s) - recombinant dna , lac operon , inducer , biopharmaceutical , escherichia coli , downstream processing , chromatography , target protein , affinity chromatography , expression vector , chemistry , fermentation , yield (engineering) , synthetic biology , transformation (genetics) , protein purification , biochemistry , microbiology and biotechnology , biology , computational biology , enzyme , gene , materials science , metallurgy
Recent advancement in fermentation technologies resulted in the increased yields of recombinant proteins of biopharmaceutical and medicinal importance. Consequently, there is an important task to develop simple and easily scalable methods that can facilitate the production of high‐quality recombinant protein. Most of the recent reports described the expression of recombinant human IL‐1 receptor antagonist (rhIL‐1Ra) in Escherichia coli using isopropyl‐β‐ d ‐thiogalacto pyranoside (IPTG), a nonmetabolizable and expensive compound, as an expression inducer. In this study, we describe the expression and one‐step purification of gallbladder‐derived rhIL‐1Ra by autoinduction in E. coli . This method includes special media that automatically induce the target protein expression from T7 promoter and allow the production of the target protein in high yield than the conventional IPTG induction method. In addition to fermentation process improvements, one‐step purification strategy is essential to make the process economical. We developed a single‐step cation exchange chromatography and obtained 300 mg/L of rhIL‐1Ra with 98% purity. Purified protein was characterized by SDS‐PAGE and Ion exchange HPLC (IEX‐HPLC). The described method can be used to scale up the production of rhIL‐1Ra and other recombinant proteins.