Reproducible preparation and effective separation of PEGylated recombinant human granulocyte colony-stimulating factor with novel “PEG-pellet” PEGylation mode and ion-exchange chromatography

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dc.contributor.authorYun, Q.
dc.contributor.authorYang, R.
dc.contributor.authorChen, T.
dc.contributor.authorBi, J.
dc.contributor.authorMa, G.
dc.contributor.authorSu, Z.
dc.date.issued2005
dc.description.abstractA novel preparation for polyethylene glycol (PEG) derivatives and chromatographic separation procedure of the PEGylated recombinant human granulocyte colony-stimulating factor (rhG-CSF) were designed to evaluate the reproducibility and scalability at large laboratory-scale level. The new “PEG-pellet” PEGylation mode was successfully applied to control the pH fluctuation during the conjugation reaction, a general problem in traditional liquid-phase conjugation mode. Moreover, two consecutive ion-exchange chromatography steps were successfully used to separate and purify the PEGylated rhG-CSF. Cation-exchange chromatography was firstly applied to separate PEGylated rhG-CSF from intact rhG-CSF, followed by anion-exchange chromatography to obtain individual PEG-rhG-CSF species (mono-, di- and tri-PEGylated rhG-CSF) and remove the excess free PEG. Furthermore, the molecular weight of individual PEGylated rhG-CSF was identified by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry and SDS-PAGE, and cell proliferation activity in vitro was detected by MTT assay using NFS-60 cell.
dc.description.urihttp://www.elsevier.com/wps/find/journaldescription.cws_home/505515/description#description
dc.identifier.citationJournal of Biotechnology, 2005; 118(1):67-74
dc.identifier.doi10.1016/j.jbiotec.2005.02.015
dc.identifier.issn0168-1656
dc.identifier.issn1873-4863
dc.identifier.orcidBi, J. [0000-0001-7056-8572]
dc.identifier.urihttp://hdl.handle.net/2440/46446
dc.language.isoen
dc.publisherElsevier Science BV
dc.source.urihttps://doi.org/10.1016/j.jbiotec.2005.02.015
dc.subjectChromatography
dc.subjectPEGylation
dc.subjectPoly(ethylene glycol)
dc.subjectProtein
dc.subjectrhG-CSF
dc.titleReproducible preparation and effective separation of PEGylated recombinant human granulocyte colony-stimulating factor with novel “PEG-pellet” PEGylation mode and ion-exchange chromatography
dc.typeJournal article
pubs.publication-statusPublished

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