File Download

There are no files associated with this item.

  Links for fulltext
     (May Require Subscription)
Supplementary

Article: High level expression, efficient purification and bioactivity assay of recombinant human platelet-derived growth factor AA dimer (PDGF-AA) from methylotrophic yeast Pichia pastoris

TitleHigh level expression, efficient purification and bioactivity assay of recombinant human platelet-derived growth factor AA dimer (PDGF-AA) from methylotrophic yeast Pichia pastoris
Authors
KeywordsGlycosylation
Recombinant protein
Human platelet-derived growth factor A
Pichia pastoris
Protein purification
Issue Date2013
Citation
Protein Expression and Purification, 2013, v. 91 n. 2, p. 221-227 How to Cite?
AbstractPlatelet-derived growth factors (PDGFs) are important biochemical mediators regulating many physiological and pathophysiological processes, including promotion of the chemotactic recruitment and proliferation of cells involved in wound repair. Previously, homodimers of rhPDGF-AA protein were purified from Escherichia coli. However, eukaryotic proteins often contain posttranslational modifications, such as glycosylation, that are required for biological functions. In this study, an efficient method was established to purify a glycosylated rhPDGF-AA dimer from P. pastoris culture media by one step CM Sepharose ion exchange chromatography yielding about 20 mg/L of over 95% highly purified rhPDGF-AA. Mass spectrometry analysis of the purified rhPDGF-AA displayed a molecular weight (MW) of 27,825.513 Da, composed of a subunit with MW of 15,042.945 Da and a subunit with MW of 12,904.374 Da. The size difference is accounted for by differential glycosylation of the monomers. Biological activity of the rhPDGF-AA was confirmed by its ability to induce NIH/3T3 cells proliferation. The experimental procedure we have developed facilitates production of an active glycosylated rhPDGF-AA in large amounts for further research and drug development. © 2013 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/189268
ISSN
2015 Impact Factor: 1.407
2015 SCImago Journal Rankings: 0.767
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorLi, Hen_US
dc.contributor.authorHui, Xen_US
dc.contributor.authorYang, Sen_US
dc.contributor.authorHu, Xen_US
dc.contributor.authorTang, Xen_US
dc.contributor.authorLi, Pen_US
dc.contributor.authorLi, Sen_US
dc.contributor.authorYang, Len_US
dc.contributor.authorJin, Sen_US
dc.contributor.authorWang, Yen_US
dc.contributor.authorXu, Aen_US
dc.contributor.authorWu, Den_US
dc.date.accessioned2013-09-17T14:31:08Z-
dc.date.available2013-09-17T14:31:08Z-
dc.date.issued2013en_US
dc.identifier.citationProtein Expression and Purification, 2013, v. 91 n. 2, p. 221-227en_US
dc.identifier.issn1046-5928-
dc.identifier.urihttp://hdl.handle.net/10722/189268-
dc.description.abstractPlatelet-derived growth factors (PDGFs) are important biochemical mediators regulating many physiological and pathophysiological processes, including promotion of the chemotactic recruitment and proliferation of cells involved in wound repair. Previously, homodimers of rhPDGF-AA protein were purified from Escherichia coli. However, eukaryotic proteins often contain posttranslational modifications, such as glycosylation, that are required for biological functions. In this study, an efficient method was established to purify a glycosylated rhPDGF-AA dimer from P. pastoris culture media by one step CM Sepharose ion exchange chromatography yielding about 20 mg/L of over 95% highly purified rhPDGF-AA. Mass spectrometry analysis of the purified rhPDGF-AA displayed a molecular weight (MW) of 27,825.513 Da, composed of a subunit with MW of 15,042.945 Da and a subunit with MW of 12,904.374 Da. The size difference is accounted for by differential glycosylation of the monomers. Biological activity of the rhPDGF-AA was confirmed by its ability to induce NIH/3T3 cells proliferation. The experimental procedure we have developed facilitates production of an active glycosylated rhPDGF-AA in large amounts for further research and drug development. © 2013 Elsevier Inc. All rights reserved.-
dc.languageengen_US
dc.relation.ispartofProtein Expression and Purificationen_US
dc.subjectGlycosylation-
dc.subjectRecombinant protein-
dc.subjectHuman platelet-derived growth factor A-
dc.subjectPichia pastoris-
dc.subjectProtein purification-
dc.titleHigh level expression, efficient purification and bioactivity assay of recombinant human platelet-derived growth factor AA dimer (PDGF-AA) from methylotrophic yeast Pichia pastorisen_US
dc.typeArticleen_US
dc.identifier.emailHui, X: hannahui@hkucc.hku.hken_US
dc.identifier.emailWang, Y: yuwanghk@hku.hken_US
dc.identifier.emailXu, A: amxu@hkucc.hku.hken_US
dc.identifier.authorityWang, Y=rp00239en_US
dc.identifier.authorityXu, A=rp00485en_US
dc.identifier.doi10.1016/j.pep.2013.08.008-
dc.identifier.pmid23978536-
dc.identifier.scopuseid_2-s2.0-84883896774-
dc.identifier.hkuros224238en_US
dc.identifier.hkuros239046-
dc.identifier.volume92en_US
dc.identifier.isiWOS:000324607100015-

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats