论文
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| 中文论文题目: | High level expression and purification of active recombinant human interleukin-8 in Pichia pastoris |
| 英文论文题目: | High level expression and purification of active recombinant human interleukin-8 in Pichia pastoris |
| 作者: | Hongbo Li , Dongye Wang , Aimin Xu , Shiwu Li , Shouguang Jin, Donghai Wu |
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| 刊物名称: | Protein Expression and Purification |
| 年: | 2009-06-21 |
| 卷: | 68 |
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| 页: | 60-64 |
| 联系作者: | Donghai Wu |
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| 影响因子: | 1.621 |
| 摘要: | 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. |
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