File Download

There are no files associated with this item.

  Links for fulltext
     (May Require Subscription)
Supplementary

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

TitleHigh level expression and purification of active recombinant human interleukin-8 in Pichia pastoris
Authors
Issue Date2009
PublisherAcademic 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?
AbstractHuman 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 Identifierhttp://hdl.handle.net/10722/59273
ISSN
2015 Impact Factor: 1.407
2015 SCImago Journal Rankings: 0.767
ISI Accession Number ID
Funding AgencyGrant Number
National Science Foundation of China30670457
30811120429
Guangzhou Administration of Science and Technology2007Z2-E4021
2005Z3-C7181
Guangzhou Economic and Technological Development District matching funds2007Ss-PO59
National 973 program of China2007CB914301
2006CB910202
2004CB720102
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 FieldValueLanguage
dc.contributor.authorLi, Hen_HK
dc.contributor.authorWang, Den_HK
dc.contributor.authorXu, Aen_HK
dc.contributor.authorLi, Sen_HK
dc.contributor.authorJin, Sen_HK
dc.contributor.authorWu, Den_HK
dc.date.accessioned2010-05-31T03:46:42Z-
dc.date.available2010-05-31T03:46:42Z-
dc.date.issued2009en_HK
dc.identifier.citationProtein Expression And Purification, 2009, v. 68 n. 1, p. 60-64en_HK
dc.identifier.issn1046-5928en_HK
dc.identifier.urihttp://hdl.handle.net/10722/59273-
dc.description.abstractHuman 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.languageengen_HK
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/yprepen_HK
dc.relation.ispartofProtein Expression and Purificationen_HK
dc.subject.meshAmmonium Sulfate - chemistryen_HK
dc.subject.meshAnimalsen_HK
dc.subject.meshCell Movementen_HK
dc.subject.meshCells, Cultureden_HK
dc.subject.meshChromatography, Affinityen_HK
dc.subject.meshCloning, Molecularen_HK
dc.subject.meshFermentationen_HK
dc.subject.meshHumansen_HK
dc.subject.meshInterleukin-8 - chemistry - genetics - metabolismen_HK
dc.subject.meshMiceen_HK
dc.subject.meshNeutrophilsen_HK
dc.subject.meshPichia - metabolismen_HK
dc.subject.meshPilot Projectsen_HK
dc.subject.meshRecombinant Proteins - chemistry - genetics - metabolismen_HK
dc.subject.meshSolubilityen_HK
dc.titleHigh level expression and purification of active recombinant human interleukin-8 in Pichia pastorisen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://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.emailXu, A:amxu@hkucc.hku.hken_HK
dc.identifier.authorityXu, A=rp00485en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.pep.2009.06.010en_HK
dc.identifier.pmid19545634-
dc.identifier.scopuseid_2-s2.0-68349141637en_HK
dc.identifier.hkuros158003en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-68349141637&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume68en_HK
dc.identifier.issue1en_HK
dc.identifier.spage60en_HK
dc.identifier.epage64en_HK
dc.identifier.isiWOS:000270927400011-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridLi, H=40261980700en_HK
dc.identifier.scopusauthoridWang, D=14051219700en_HK
dc.identifier.scopusauthoridXu, A=7202655409en_HK
dc.identifier.scopusauthoridLi, S=26039280900en_HK
dc.identifier.scopusauthoridJin, S=7401822323en_HK
dc.identifier.scopusauthoridWu, D=7404297751en_HK
dc.identifier.citeulike5458213-

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats