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Integrated Extraction-Preservation Strategies for RNA Using Biobased Ionic Liquids

2019Journal article Restricted access
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dc.contributor.authorQuental, Maria V.
dc.contributor.authorPedro, Augusto
dc.contributor.authorPereira, Patrícia
dc.contributor.authorSharma, Mukesh
dc.contributor.authorQueiroz, João
dc.contributor.authorCoutinho, João A.P.
dc.contributor.authorSousa, Fani
dc.contributor.authorFreire, Mara G.
dc.date.accessioned2020-01-09T17:18:40Z
dc.date.available2020-01-09T17:18:40Z
dc.date.issued2019
dc.description.abstractThe ubiquitous instability of RNA along with issues associated with its purity degree have been preventing its widespread use as low-cost biotherapeutics. On the basis of the well-known capacity of amino acids to specifically interact with RNA when used as chromatographic ligands, a set of amino-acid-based ionic liquids (AA-ILs) was herein investigated, both to act as preservation media and as phase-forming agents of aqueous biphasic systems (ABS). This set of strategies was combined with the goal of developing integrated extraction-preservation platforms. AA-ILs comprising the cholinium cation and anions derived from l-lysine ([Ch][Lys]), l-arginine ([Ch][Arg]), l-glutamic acid ([Ch][Glu]), and dl-aspartic acid ([Ch][Asp]) were studied. It is shown that the stability of RNA is preserved in aqueous solutions of the studied AA-ILs, even in the presence of ribonucleases (RNases). Furthermore, almost all the investigated AA-ILs display no cytotoxicity onto two distinct human cell lines. After identifying the most promising ILs, ABS formed by AA-ILs and polypropylene glycol with a molecular weight of 400 g mol–1 (PPG 400) were investigated as extraction and purification platforms for RNA. Both with pure RNA and bacterial lysate samples, RNA is successfully extracted to the IL-rich phase without compromising its integrity and stability. On the basis of these results, the integrated extraction-preservation process for RNA is finally demonstrated. RNA is initially extracted from the bacterial lysate sample using ABS, after which the IL-rich phase can be used as the preservation medium of RNA up to its use. RNA can be then recovered from the IL-rich phase by ethanol precipitation, and the ABS phase-forming components recovered and reused. Although improvements in the purity level of RNA are still required, the approach here reported represents a step forward in the development of sustainable processes to overcome the critical demand of high-quality/high-purity RNA to be used as biotherapeutics.pt_PT
dc.description.sponsorshipEuropean Research Council (ERC) for the Starting Grant ERC-2013-StG-337753pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1021/acssuschemeng.9b00688pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/8185
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherACS Publicationspt_PT
dc.relationPurification of antibodies Immunoglobulin Y, IgY using aqueous biphasic systems composed of novel ionic liquids with buffering characteristics
dc.relationCICECO-Aveiro Institute of Materials
dc.relationDetermination of the architecture and the RNA degradation strategy of Ribonuclease R: implications for pathogen control.
dc.relation.publisherversionhttps://pubs.acs.org/doi/10.1021/acssuschemeng.9b00688pt_PT
dc.subjectRibonucleic acidpt_PT
dc.subjectAmino-acid-based ionic liquidspt_PT
dc.subjectAqueous biphasic systemspt_PT
dc.subjectIntegrated extraction-preservation processpt_PT
dc.titleIntegrated Extraction-Preservation Strategies for RNA Using Biobased Ionic Liquidspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitlePurification of antibodies Immunoglobulin Y, IgY using aqueous biphasic systems composed of novel ionic liquids with buffering characteristics
oaire.awardTitleCICECO-Aveiro Institute of Materials
oaire.awardTitleDetermination of the architecture and the RNA degradation strategy of Ribonuclease R: implications for pathogen control.
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBD%2F109765%2F2015/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/UID%2FMulti%2F00709%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID%2FCTM%2F50011%2F2019/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FBIA-MIC%2F1399%2F2014/PT
oaire.citation.endPage9448pt_PT
oaire.citation.issue10pt_PT
oaire.citation.startPage9439pt_PT
oaire.citation.titleACS Sustainable Chemistry and Engineeringpt_PT
oaire.citation.volume7pt_PT
oaire.fundingStream5876
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream3599-PPCDT
person.familyNameQuaresma Henriques Pedro
person.familyNamePereira
person.familyNameQueiroz
person.familyNameCoutinho
person.familyNameSousa
person.familyNameFreire
person.givenNameAugusto
person.givenNamePatrícia
person.givenNameJoão
person.givenNameJoao
person.givenNameFani
person.givenNameMara
person.identifierhttps://scholar.google.com/citations?user=oer44UoAAAAJ&hl=pt-PT&oi=ao
person.identifier308549
person.identifier.ciencia-id7111-B2BE-336D
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person.identifier.ciencia-id991D-2E13-A840
person.identifier.ciencia-id6718-1145-692F
person.identifier.orcid0000-0001-8194-6022
person.identifier.orcid0000-0002-5665-5271
person.identifier.orcid0000-0002-3096-8325
person.identifier.orcid0000-0002-3841-743X
person.identifier.orcid0000-0001-9996-2194
person.identifier.orcid0000-0001-8895-0614
person.identifier.ridL-3104-2014
person.identifier.ridA-2014-2017
person.identifier.ridB-8458-2008
person.identifier.scopus-author-id54080426100
person.identifier.scopus-author-id56111844900
person.identifier.scopus-author-id7003705645
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person.identifier.scopus-author-id7005110268
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project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
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
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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