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- Publisher Website: 10.1016/j.pep.2009.06.010
- Scopus: eid_2-s2.0-68349141637
- PMID: 19545634
- WOS: WOS:000270927400011
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Article: High level expression and purification of active recombinant human interleukin-8 in Pichia pastoris
Title | High level expression and purification of active recombinant human interleukin-8 in Pichia pastoris | ||||||||||
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Authors | |||||||||||
Keywords | Expression and purification Interleukin-8 Pichia pastoris Recombinant protein | ||||||||||
Issue Date | 2009 | ||||||||||
Publisher | Academic Press. The Journal's web site is located at http://www.elsevier.com/locate/yprep | ||||||||||
Citation | Protein Expression And Purification, 2009, v. 68 n. 1, p. 60-64 How to Cite? | ||||||||||
Abstract | 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. © 2009 Elsevier Inc. | ||||||||||
Persistent Identifier | http://hdl.handle.net/10722/59273 | ||||||||||
ISSN | 2023 Impact Factor: 1.4 2023 SCImago Journal Rankings: 0.383 | ||||||||||
ISI Accession Number ID |
Funding Information: This work was supported in part by Funds from the National Science Foundation of China (30670457 and 30811120429), Guangzhou Administration of Science and Technology (2007Z2-E4021 and 2005Z3-C7181), Guangzhou Economic and Technological Development District matching funds (2007Ss-PO59), and the National 973 program of China (2007CB914301, 2006CB910202, and 2004CB720102). | ||||||||||
References |
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Li, H | en_HK |
dc.contributor.author | Wang, D | en_HK |
dc.contributor.author | Xu, A | en_HK |
dc.contributor.author | Li, S | en_HK |
dc.contributor.author | Jin, S | en_HK |
dc.contributor.author | Wu, D | en_HK |
dc.date.accessioned | 2010-05-31T03:46:42Z | - |
dc.date.available | 2010-05-31T03:46:42Z | - |
dc.date.issued | 2009 | en_HK |
dc.identifier.citation | Protein Expression And Purification, 2009, v. 68 n. 1, p. 60-64 | en_HK |
dc.identifier.issn | 1046-5928 | en_HK |
dc.identifier.uri | http://hdl.handle.net/10722/59273 | - |
dc.description.abstract | 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. © 2009 Elsevier Inc. | en_HK |
dc.language | eng | en_HK |
dc.publisher | Academic Press. The Journal's web site is located at http://www.elsevier.com/locate/yprep | en_HK |
dc.relation.ispartof | Protein Expression and Purification | en_HK |
dc.subject | Expression and purification | - |
dc.subject | Interleukin-8 | - |
dc.subject | Pichia pastoris | - |
dc.subject | Recombinant protein | - |
dc.subject.mesh | Ammonium Sulfate - chemistry | en_HK |
dc.subject.mesh | Animals | en_HK |
dc.subject.mesh | Cell Movement | en_HK |
dc.subject.mesh | Cells, Cultured | en_HK |
dc.subject.mesh | Chromatography, Affinity | en_HK |
dc.subject.mesh | Cloning, Molecular | en_HK |
dc.subject.mesh | Fermentation | en_HK |
dc.subject.mesh | Humans | en_HK |
dc.subject.mesh | Interleukin-8 - chemistry - genetics - metabolism | en_HK |
dc.subject.mesh | Mice | en_HK |
dc.subject.mesh | Neutrophils | en_HK |
dc.subject.mesh | Pichia - metabolism | en_HK |
dc.subject.mesh | Pilot Projects | en_HK |
dc.subject.mesh | Recombinant Proteins - chemistry - genetics - metabolism | en_HK |
dc.subject.mesh | Solubility | en_HK |
dc.title | High level expression and purification of active recombinant human interleukin-8 in Pichia pastoris | en_HK |
dc.type | Article | en_HK |
dc.identifier.openurl | http://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1046-5928&volume=68&spage=60&epage=64&date=2009&atitle=High+level+expression+and+purification+of+active+recombinant+human+interleukin-8+in+Pichia+pastoris. | en_HK |
dc.identifier.email | Xu, A:amxu@hkucc.hku.hk | en_HK |
dc.identifier.authority | Xu, A=rp00485 | en_HK |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1016/j.pep.2009.06.010 | en_HK |
dc.identifier.pmid | 19545634 | - |
dc.identifier.scopus | eid_2-s2.0-68349141637 | en_HK |
dc.identifier.hkuros | 158003 | en_HK |
dc.relation.references | http://www.scopus.com/mlt/select.url?eid=2-s2.0-68349141637&selection=ref&src=s&origin=recordpage | en_HK |
dc.identifier.volume | 68 | en_HK |
dc.identifier.issue | 1 | en_HK |
dc.identifier.spage | 60 | en_HK |
dc.identifier.epage | 64 | en_HK |
dc.identifier.isi | WOS:000270927400011 | - |
dc.publisher.place | United States | en_HK |
dc.identifier.scopusauthorid | Li, H=40261980700 | en_HK |
dc.identifier.scopusauthorid | Wang, D=14051219700 | en_HK |
dc.identifier.scopusauthorid | Xu, A=7202655409 | en_HK |
dc.identifier.scopusauthorid | Li, S=26039280900 | en_HK |
dc.identifier.scopusauthorid | Jin, S=7401822323 | en_HK |
dc.identifier.scopusauthorid | Wu, D=7404297751 | en_HK |
dc.identifier.citeulike | 5458213 | - |
dc.identifier.issnl | 1046-5928 | - |