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Prediction of retention time of cutinases tagged with hydrophobic peptides in hydrophobic interaction chromatography

2007-06-22Journal article Restricted access
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dc.contributor.authorLienqueo, M. E.
dc.contributor.authorSalazar, O.
dc.contributor.authorHenriquez, K.
dc.contributor.authorCalado, Cecília
dc.contributor.authorFonseca, L. P.
dc.contributor.authorCabral, J. M. S.
dc.date.accessioned2020-09-22T13:20:38Z
dc.date.available2020-09-22T13:20:38Z
dc.date.issued2007-06-22
dc.description.abstractHydrophobic interaction chromatography (HIC) is an important technique for protein purification, which exploits the separation of proteins based on hydrophobic interactions between the stationary phase ligands and hydrophobic regions on the protein surface. One way of enhancing the purification efficiency by HIC is the addition of short sequences of peptide tags to the target protein by genetic engineering, which could reduce the need for extra and expensive chromatographic steps. In the present work, a methodology for predicting retention times of cutinases tagged with hydrophobic peptides in HIC is presented. Cutinase from Fusarium solani pisi fused to tryptophan–proline (WP) tags, namely (WP)2 and (WP)4, and produced in Saccharomyces cerevisiae strains, were used as model proteins. From the simulations, the methodology based on tagged hydrophobic definition proposed by Simeonidis et al. (Φtagged), associated to a quadratic model for predicting dimensionless retention times, showed small differences (RMSE < 0.022) between observed and estimated retention times. The difference between observed and calculated retention times being lower than 2.0% (RMSE < 0.022) for the two tagged cutinases at three different stationary phases, except for the case of cut_(wp)2 in octyl sepharose–2 M ammonium sulphate. Therefore, we consider that the proposed strategy, based on tagged surface hydrophobicity, allows prediction of acceptable retention times of cutinases tagged with hydrophobic peptides in HIC.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationLIENQUEO, M. E.; [et al] – Prediction of retention time of cutinases tagged with hydrophobic peptides in hydrophobic interaction chromatography. Journal of Chromatography A. ISSN 0021-9673. Vol. 1154, N.º 1-2 (2007), pp. 460-463pt_PT
dc.identifier.doi10.1016/j.chroma.2007.03.088pt_PT
dc.identifier.issn0021-9673
dc.identifier.urihttp://hdl.handle.net/10400.21/12250
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relation.publisherversionhttps://reader.elsevier.com/reader/sd/pii/S0021967307006152?token=F3FD1FACDDE0B9AE0D67E83F2C7F0C0A4A73B8E630701A21729B5C07ED3113419F9F4CACB56844FDC90CE0E2DE3E23D4pt_PT
dc.subjectHydrophobic interaction chromatographypt_PT
dc.subjectProtein hydrophobicitypt_PT
dc.subjectRetention time predictionpt_PT
dc.subjectHydrophobic peptide tagspt_PT
dc.titlePrediction of retention time of cutinases tagged with hydrophobic peptides in hydrophobic interaction chromatographypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBPD%2F11626%2F2002/PT
oaire.citation.endPage463pt_PT
oaire.citation.issue1-2pt_PT
oaire.citation.startPage460pt_PT
oaire.citation.titleJournal of Chromatography Apt_PT
oaire.citation.volume1154pt_PT
oaire.fundingStreamSFRH
person.familyNameCalado
person.givenNameCecília
person.identifier130332
person.identifier.ciencia-id9418-E320-3177
person.identifier.orcid0000-0002-5264-9755
person.identifier.ridE-2102-2014
person.identifier.scopus-author-id6603163260
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicatione8577257-c64c-4481-9b2b-940fedb360cc
relation.isAuthorOfPublication.latestForDiscoverye8577257-c64c-4481-9b2b-940fedb360cc
relation.isProjectOfPublication4adab492-5d13-4951-9a33-79b601598702
relation.isProjectOfPublication.latestForDiscovery4adab492-5d13-4951-9a33-79b601598702

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