High level expression and purification of active recombinant human interleukin-8 in Pichia pastoris

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• 作者：Hongbo Li ; Dongye Wang ; Aimin Xu ; Shiwu Li ; Shouguang Jin ; Donghai Wu
• 关键词：Interleukin-8 ; Recombinant protein ; Pichia pastoris ; Expression and purification
• 刊名：Protein Expression and Purification
• 出版年：2009
• 出版时间：November 2009
• 年：2009
• 卷：68
• 期：1
• 页码：60-64
• 全文大小：747 K

文摘

Human interleukin-8 (hIL-8) is a member of interleukin family which functions as a chemotactic factor as well as an angiogenesis mediator. Previously, a study reported that hIL-8 could be purified from inclusion bodies using a prokaryotic expression system, however, the required re-naturation step limits the recovery of fully active protein. In this study, soluble recombinant hIL-8 was expressed as a secreted protein at high level in Pichia pastoris under the control of AOX1 (alcohol oxidase 1) promoter. A simple purification strategy was established to recover rhIL-8 from the fermentation supernatant. The process includes precipitation with 80 % saturation ammonium sulfate and CM Sepharose ion exchange chromatography, yielding 30 mg/L purified rhIL-8 at over 95 % purity. The obtained rhIL-8 displays high specific activity, stimulating the migration of mouse neutrophils at concentrations as low as 0.25 ng/mL. Our results demonstrate that P. pastoris expression system is an efficient tool for large-scale manufacture of active recombinant hIL-8 for various applications.