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Purification of pre-miR-29 by arginine-affinity chromatography

2014-01-19Journal article Restricted access
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dc.contributor.authorPereira, Patrícia
dc.contributor.authorSousa, Ângela
dc.contributor.authorQueiroz, João
dc.contributor.authorCorreia, Ilídio
dc.contributor.authorFigueiras, Ana
dc.contributor.authorSousa, Fani
dc.date.accessioned2018-03-20T14:54:57Z
dc.date.available2018-03-20T14:54:57Z
dc.date.issued2014-01-19
dc.description.abstractRecently, differential expression of microRNAs, in patients with Alzheimer's disease (AD) suggests that they might have key regulatory roles in this neurodegenerative disease. Taking into account this fact, several studies demonstrated that the miR-29 is significantly decreased in AD patients, also displaying abnormally high levels of β-site APP-cleaving enzyme 1. Thus, RNA biochemical or structural studies often require a RNA sample that is chemically pure and biologically active. The present work describes a new affinity chromatography method using an arginine support to specifically purify pre-miR-29 from other Rhodovulum sulfidophilum small RNA species. Nevertheless, in order to achieve higher efficiency and selectivity, it is essential to characterize the behavior of pre-miR-29 binding/elution. Thus, three different strategies based on increased sodium chloride (280–500 mM), arginine (25 mM) or decreased ammonium sulfate (2–0.1 M) stepwise gradients are described to purify pre-miR-29. In this way, it was proved that well-defined binding/elution conditions are crucial to enhance the purification performance. As a matter of fact, by employing elution strategies using sodium chloride or arginine, an improvement in the final pre-miR-29 yields (96.5 and 56.7%, respectively) was obtained. Moreover, the quality control analysis revealed high integrity in pre-miR-29 preparations as well as high purity (90 and 98%, respectively), demonstrated by the scarce detection of proteins. This improved method takes advantage of its simplicity, significant cost reduction, due to the elimination of some complex operations, and speed for large-scale purification of pre-miRNAs suitable for biochemical and structural studies.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationPereira, P., Sousa, A., Queiroz, J.A., Correia, I.J., Figueiras, A. e Sousa, F. (2014) "Purification of pre-miR-29 by arginine-affinity chromatography", Journal of Chromatography B, Vol. 951-952, pp. 16-23pt_PT
dc.identifier.doi10.1016/j.jchromb.2014.01.020pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/4662
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relationIsolation and Purification of Plasmid DNA for Cancer Therapy
dc.relationStrategic Project - UI 709 - 2011-2012
dc.relationMIRNA-29 BIOSEPARATION AND TARGET DELIVERY STRATEGIES FOR ALZHEIMER DISEASE
dc.relationPURIFICATION OF HPV-16 E6/E7 DNA VACCINE BY MONOLITHIC CHROMATOGRAPHY USING AMINO ACIDS AND DERIVATIVES AS AFFINITY LIGANDS FOR PHARMACEUTICAL APPLICATIONS
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S1570023214000257?via%3Dihubpt_PT
dc.subjectAlzheimer's diseasept_PT
dc.subjectArginine-affinity chromatographypt_PT
dc.subjectMultiple interactionspt_PT
dc.subjectPre-miR-29pt_PT
dc.subjectRhodovulum sulfidophilumpt_PT
dc.subjectRNA purificationpt_PT
dc.titlePurification of pre-miR-29 by arginine-affinity chromatographypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleIsolation and Purification of Plasmid DNA for Cancer Therapy
oaire.awardTitleStrategic Project - UI 709 - 2011-2012
oaire.awardTitleMIRNA-29 BIOSEPARATION AND TARGET DELIVERY STRATEGIES FOR ALZHEIMER DISEASE
oaire.awardTitlePURIFICATION OF HPV-16 E6/E7 DNA VACCINE BY MONOLITHIC CHROMATOGRAPHY USING AMINO ACIDS AND DERIVATIVES AS AFFINITY LIGANDS FOR PHARMACEUTICAL APPLICATIONS
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876-PPCDTI/PTDC%2FEBB-BIO%2F114320%2F2009/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6820 - DCRRNI ID/PEst-C%2FSAU%2FUI0709%2F2011/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/FARH/SFRH%2FBD%2F81914%2F2011/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876-PPCDTI/EXPL%2FBBB-BIO%2F1056%2F2012/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/FARH/SFRH%2FBPD%2F79106%2F2011/PT
oaire.citation.endPage23pt_PT
oaire.citation.startPage16pt_PT
oaire.citation.titleJournal of Chromatography Bpt_PT
oaire.citation.volume951-952pt_PT
oaire.fundingStream5876-PPCDTI
oaire.fundingStream6820 - DCRRNI ID
oaire.fundingStreamFARH
oaire.fundingStream5876-PPCDTI
oaire.fundingStreamFARH
person.familyNamePereira
person.familyNameSousa
person.familyNameQueiroz
person.familyNameJoaquim Sobreira Correia
person.familyNameFigueiras
person.familyNameSousa
person.givenNamePatrícia
person.givenNameÂngela
person.givenNameJoão
person.givenNameIlídio
person.givenNameAna
person.givenNameFani
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person.identifier.ridL-3104-2014
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project.funder.identifierhttp://doi.org/10.13039/501100001871
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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
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.embargofctCopyright cedido à editora no momento da publicaçãopt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
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