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Separation of human immunoglobulin G subclasses on a protein A monolith column

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dc.contributor.authorLeblebici, Pelin
dc.contributor.authorLeblebici, Mumin Enis
dc.contributor.authorFerreira-da-Silva, Frederico
dc.contributor.authorRodrigues, Alírio
dc.contributor.authorPais, Luís S.
dc.date.accessioned2014-07-14T09:28:52Z
dc.date.available2014-07-14T09:28:52Z
dc.date.issued2014
dc.description.abstractMonolithic columns have attracted significant attention for the purification of large biomolecules. In thepresent study, a step gradient elution method was evaluated for the separation of human immunoglobulinG (hIgG) into its subclasses on CIM (convective interaction media) r-protein A (recombinant protein A)monolithic column. hIgG was loaded onto the column and bound protein was eluted with a pH gra-dient. The subclass content of the eluted fractions was analyzed by enzyme-linked immunosorbentassay (ELISA). Results showed that separation of IgG3 from the other three subclasses can be success-fully achieved with high selectivity (100%) and throughput on monolithic media. It was also revealedthat enriched fractions of IgG1 and IgG2 could be obtained from purified hIgG in a 28 min long chro-matographic run. Three fractions with high IgG1 content (89.1%, 94.3% and 88.8%) were recovered.Furthermore, IgG2 was enriched to 64% successfully. A rapid step gradient elution scheme without anyadditives in buffers was proven to obtain enriched preparations of the two important subclasses withhigh throughput. The separation time can be reduced even more by increasing the flow rate without anyloss in selectivity, which will be beneficial in industrial scale applications.por
dc.identifier.citationLeblebici, P.; Leblebici, M.E.; Ferreira-da-Silva, F.; Rodrigues, A.E.; Pais, L.S. (2014). Separation of human immunoglobulin G subclasses on a protein A monolith column. Journal of chromatography B. ISSN 1570-0232. 962, p. 89-93por
dc.identifier.doi10.1016/j.jchromb.2014.05.029
dc.identifier.issn1570-0232
dc.identifier.urihttp://hdl.handle.net/10198/9898
dc.language.isoengpor
dc.peerreviewedyespor
dc.relationAdTechP2 - Advanced Separation Technologies for the Protein Purification
dc.subjectImmunoglobulin G subclassespor
dc.subjectMonolithpor
dc.subjectSeparationpor
dc.subjectProtein A chromatographypor
dc.titleSeparation of human immunoglobulin G subclasses on a protein A monolith columnpor
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleAdTechP2 - Advanced Separation Technologies for the Protein Purification
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876-PPCDTI/PTDC%2FEQU-EQU%2F113310%2F2009/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/COMPETE/PEst-C%2FEQB%2FLA0020%2F2013/PT
oaire.citation.endPage93por
oaire.citation.startPage89por
oaire.citation.titleJournal of chromatography. B.por
oaire.citation.volume962por
oaire.fundingStream5876-PPCDTI
oaire.fundingStreamCOMPETE
person.familyNamePais
person.givenNameLuís S.
person.identifier.ciencia-id421E-0CCD-A84F
person.identifier.orcid0000-0002-3000-2862
person.identifier.scopus-author-id6603930478
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor
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relation.isAuthorOfPublication.latestForDiscoveryc6d25534-8487-45b1-90af-d350f4e3ac55
relation.isProjectOfPublicationfded14df-10b9-4a14-a837-6e36057ee0e6
relation.isProjectOfPublicatione843830d-c7d3-427f-9155-f20b821965fc
relation.isProjectOfPublication.latestForDiscoveryfded14df-10b9-4a14-a837-6e36057ee0e6

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