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Article: Expression, purification, and functional characterization of recombinant human interleukin-7

TitleExpression, purification, and functional characterization of recombinant human interleukin-7
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. 63 n. 1, p. 1-4 How to Cite?
AbstractHuman interleukin-7 (IL-7) is a member of the interleukin family. Numerous studies have demonstrated IL-7's effect on B- and T-cell development as well as its potential in various clinical applications. Previously, a study reported that IL-7 could be purified from inclusion bodies using a prokaryotic system, however, the required refolding step limits the recovery rate. This study was designed to produce a bioactive recombinant human IL-7 (rhIL-7) in a eukaryotic expression system in order to obtain higher yields of the protein with simpler purification steps. We cloned human IL-7 cDNA and successfully expressed active recombinant protein in yeast using the Pichia pastoris expression system. A simple purification strategy was established to purify the rhIL-7 from the fermentation supernatant, yielding 35 mg/L at 95% purity by the use of a common SP Sepharose FF cation-exchange chromatography. Functional analysis of the purified rhIL-7 by the pre-B cell MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) proliferation assay demonstrated a specific activity comparable to commercial sources. These results suggest that purification of rhIL-7 from yeast provides a sound strategy for large-scale production of the rhIL-7 for clinical applications as well as basic researches. © 2008 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/59274
ISSN
2015 Impact Factor: 1.407
2015 SCImago Journal Rankings: 0.767
ISI Accession Number ID
Funding AgencyGrant Number
National Science Foundation of China30670457
Guangzhou Administration of Science and Technology2007Z2-E4021
2005Z3-C7181
Guangzhou Economic and Technological Development District matching funds2007Ss-P059
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), Guangzhou Administration of Science and Technology (2007Z2-E4021 and 2005Z3-C7181), Guangzhou Economic and Technological Development District matching funds (2007Ss-P059) and the National 973 program of China (2007CB914301, 2006CB910202 and 2004CB720102).

References

 

DC FieldValueLanguage
dc.contributor.authorLuo, Yen_HK
dc.contributor.authorKong, Xen_HK
dc.contributor.authorXu, Aen_HK
dc.contributor.authorJin, Sen_HK
dc.contributor.authorWu, Den_HK
dc.date.accessioned2010-05-31T03:46:43Z-
dc.date.available2010-05-31T03:46:43Z-
dc.date.issued2009en_HK
dc.identifier.citationProtein Expression And Purification, 2009, v. 63 n. 1, p. 1-4en_HK
dc.identifier.issn1046-5928en_HK
dc.identifier.urihttp://hdl.handle.net/10722/59274-
dc.description.abstractHuman interleukin-7 (IL-7) is a member of the interleukin family. Numerous studies have demonstrated IL-7's effect on B- and T-cell development as well as its potential in various clinical applications. Previously, a study reported that IL-7 could be purified from inclusion bodies using a prokaryotic system, however, the required refolding step limits the recovery rate. This study was designed to produce a bioactive recombinant human IL-7 (rhIL-7) in a eukaryotic expression system in order to obtain higher yields of the protein with simpler purification steps. We cloned human IL-7 cDNA and successfully expressed active recombinant protein in yeast using the Pichia pastoris expression system. A simple purification strategy was established to purify the rhIL-7 from the fermentation supernatant, yielding 35 mg/L at 95% purity by the use of a common SP Sepharose FF cation-exchange chromatography. Functional analysis of the purified rhIL-7 by the pre-B cell MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) proliferation assay demonstrated a specific activity comparable to commercial sources. These results suggest that purification of rhIL-7 from yeast provides a sound strategy for large-scale production of the rhIL-7 for clinical applications as well as basic researches. © 2008 Elsevier Inc. All rights reserved.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.meshBioreactorsen_HK
dc.subject.meshBlotting, Westernen_HK
dc.subject.meshChromatography, Ion Exchangeen_HK
dc.subject.meshCloning, Molecularen_HK
dc.subject.meshFermentationen_HK
dc.subject.meshHumansen_HK
dc.subject.meshInterleukin-7 - biosynthesis - genetics - isolation & purification - pharmacologyen_HK
dc.subject.meshPichia - genetics - metabolismen_HK
dc.subject.meshProtein Renaturationen_HK
dc.subject.meshRecombinant Proteins - biosynthesis - genetics - isolation & purification - pharmacologyen_HK
dc.subject.meshSepharoseen_HK
dc.subject.meshTetrazolium Saltsen_HK
dc.subject.meshThiazolesen_HK
dc.subject.meshTransformation, Geneticen_HK
dc.titleExpression, purification, and functional characterization of recombinant human interleukin-7en_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1046-5928&volume=63&spage=1&epage=4&date=2009&atitle=Expression,+purification,+and+functional+characterization+of+recombinant+human+interleukin-7.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.2008.08.009en_HK
dc.identifier.pmid18805490-
dc.identifier.scopuseid_2-s2.0-54849426172en_HK
dc.identifier.hkuros158001en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-54849426172&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume63en_HK
dc.identifier.issue1en_HK
dc.identifier.spage1en_HK
dc.identifier.epage4en_HK
dc.identifier.isiWOS:000261035100001-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridLuo, Y=55187972100en_HK
dc.identifier.scopusauthoridKong, X=7202794607en_HK
dc.identifier.scopusauthoridXu, A=7202655409en_HK
dc.identifier.scopusauthoridJin, S=7401822323en_HK
dc.identifier.scopusauthoridWu, D=7404297751en_HK

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