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Arginine and di-arginine ligands for plasmid DNA purification using negative chromatography

2018Journal article Restricted access
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dc.contributor.authorCardoso, Sara
dc.contributor.authorFilho, Pedro De Alcântara Pessôa
dc.contributor.authorSousa, Fani
dc.contributor.authorAzzoni, Adriano
dc.date.accessioned2020-01-09T17:04:23Z
dc.date.available2020-01-09T17:04:23Z
dc.date.issued2018
dc.description.abstractThe increasing number of applications requiring highly purified plasmid DNA (pDNA) generates a corresponding need for simple, scalable, and cost-effective purification processes. Due to the pDNA large size and complex shape, the use of commercial chromatographic beads often results in poor yields and low binding capacities when operated in a positive mode. An alternative to overcome this limitation is the design of chromatographic ligand-resin systems able to efficiently operate in negative mode, where host impurities (especially low molecular weight RNA) are efficiently captured and separated from the target pDNA. In this work, arginine amino acid and di-arginine peptide (arginine-arginine) were immobilized in agarose resins and evaluated for negative chromatographic purification of pDNA from bacterial cell lysates. The results showed that RNA was preferentially bound to the ligands, interfering with the binding of pDNA. The amount of plasmid processed per column volume by arginine and di-arginine, under negative mode, was substantially larger comparing with the conventional positive mode, resulting in pDNA recoveries up to 99%, with a considerable reduction of host impurities. This study shows that negative mode chromatography using arginine-based ligands poses as an interesting alternative for intermediate and polishing pDNA purification operations, with considerable economic and environmental advantages.pt_PT
dc.description.sponsorshipThis work was supported by CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico (grants 444412/2014-0, 304906/2014-0 and 307739/2015-5), and FAPESP - Fundação de Amparo à Pesquisa do Estado de São Paulo (grant 2013/23780-1). The authors acknowledges to Professor Miguel Prazeres, from Instituto Superior Técnico, Lisboa, Portugal, for the kindly donation of the E. coli DH10b harboring the pVAX1-GFP plasmid.
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1016/j.seppur.2018.03.066pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/8184
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S1383586617339059pt_PT
dc.subjectPlasmid DNA purificationpt_PT
dc.subjectPlasmid DNA purificationpt_PT
dc.subjectArgininept_PT
dc.subjectArgininept_PT
dc.subjectAgarose resinpt_PT
dc.titleArginine and di-arginine ligands for plasmid DNA purification using negative chromatographypt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage289pt_PT
oaire.citation.startPage281pt_PT
oaire.citation.titleSeparation and Purification Technologypt_PT
oaire.citation.volume202pt_PT
person.familyNamePessôa Filho
person.familyNameSousa
person.familyNameAzzoni
person.givenNamePedro de Alcântara
person.givenNameFani
person.givenNameAdriano
person.identifier.ciencia-id991D-2E13-A840
person.identifier.orcid0000-0003-4315-7238
person.identifier.orcid0000-0001-9996-2194
person.identifier.orcid0000-0003-0696-4663
person.identifier.ridA-2014-2017
person.identifier.scopus-author-id6603339745
person.identifier.scopus-author-id7005110268
person.identifier.scopus-author-id7003926883
rcaap.embargofctCopyright cedido à editora no momento da publicaçãopt_PT
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication90e8e7bb-dad1-4e58-a45d-73729819c973
relation.isAuthorOfPublication2935de97-fc90-4d38-9505-4caa364f8a10
relation.isAuthorOfPublication8f8ddda3-7e25-443a-be1a-d5371e040f51
relation.isAuthorOfPublication.latestForDiscovery90e8e7bb-dad1-4e58-a45d-73729819c973

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