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Improved Minicircle DNA Biosynthesis for Gene Therapy Applications

2014-04-01Journal article Restricted access
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dc.contributor.authorGaspar, V. M.
dc.contributor.authorMaia, C J
dc.contributor.authorQueiroz, João
dc.contributor.authorPichon, Chantal
dc.contributor.authorCorreia, Ilídio
dc.contributor.authorSousa, Fani
dc.date.accessioned2018-03-20T11:29:08Z
dc.date.available2018-03-20T11:29:08Z
dc.date.issued2014-04-01
dc.description.abstractMinicircular DNA (mcDNA) biopharmaceuticals have recently risen as a valuable alternative for the development of a next generation of bioactive therapeutics because they are more efficient and safer than standard plasmid DNA (pDNA). To date, the relatively insufficient knowledge regarding mcDNA biosynthesis is currently hindering its manufacture in suitable amounts for clinical trial evaluations. Addressing this limitation is therefore mandatory to bring forth the full therapeutic potential of this cutting-edge technology. Herein, we describe for the first time new processing parameters that improve the overall yield of mcDNA obtained from bacterial fermentations. We provide details for further in-line monitoring and optimization in view of the current good manufacturing guidelines. Our results show that by rising growth temperature to 42°C, an increase in the overall minicircle producer plasmid yield is attained, while biomass amounts are reduced. Moreover, by monitoring in real time the dynamic recombination of parental plasmids to mcDNA, we found that this event is more efficient at specific time points, regardless of the growth temperature and inductor concentration used. These are important findings since mcDNA can be recovered with higher yields at these determined key stages. Indeed, the manipulation of these parameters resulted in a 2.21-fold increase in mcDNA production compared with the established growth temperatures for this technology. Overall, our findings highlight that to achieve maximum productivity while attaining pharmaceutical-grade mcDNA preparations, process design and biosynthesis optimization must take into account key parameters such as temperature, inductor concentration, and recovery time.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationGaspar, V.M., Maia, C.J., Queiroz, J.A., Pichon, C., Correia, I.J. e Sousa, F. (2014) "Improved minicircle dna biosynthesis for gene therapy applications", Human Gene Therapy Methods, Vol. 25(2), pp. 93-105pt_PT
dc.identifier.doi10.1089/hgtb.2013.020pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/4654
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherMary Ann Liebertpt_PT
dc.relationIsolation and Purification of Plasmid DNA for Cancer Therapy
dc.relationStrategic Project - UI 709 - 2011-2012
dc.relationBIOSYNTHESIS AND PURIFICATION OF MINICIRCLE DNA FOR APPLICATION IN DIABETES CELL-SPECIFIC GENE THERAPY
dc.relation.publisherversionhttps://www.liebertpub.com/doi/abs/10.1089/hgtb.2013.020?url_ver=Z39.88-2003&rfr_id=ori%3Arid%3Acrossref.org&rfr_dat=cr_pub%3Dpubmedpt_PT
dc.subjectMinicircle DNApt_PT
dc.subjectBiosynthesispt_PT
dc.subjectGene Therapypt_PT
dc.titleImproved Minicircle DNA Biosynthesis for Gene Therapy Applicationspt_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.awardTitleBIOSYNTHESIS AND PURIFICATION OF MINICIRCLE DNA FOR APPLICATION IN DIABETES CELL-SPECIFIC GENE THERAPY
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FEME-TME%2F103375%2F2008/PT
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%2F80402%2F2011/PT
oaire.citation.endPage105pt_PT
oaire.citation.startPage93pt_PT
oaire.citation.titleHuman Gene Therapy Methodspt_PT
oaire.citation.volume25pt_PT
oaire.fundingStream3599-PPCDT
oaire.fundingStream5876-PPCDTI
oaire.fundingStream6820 - DCRRNI ID
oaire.fundingStreamFARH
person.familyNameGaspar
person.familyNameMaia Baptista
person.familyNameQueiroz
person.familyNameCorreia
person.familyNameSousa
person.givenNameVítor
person.givenNameCláudio Jorge
person.givenNameJoão
person.givenNameIlídio Joaquim Sobreira
person.givenNameFani
person.identifierhttps://scholar.google.pt/citations?user=hFKOd_QAAAAJ&hl=pt-PT&oi=ao
person.identifierUMbJ1KMAAAAJ
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person.identifier.ciencia-id991D-2E13-A840
person.identifier.orcid0000-0002-0372-2493
person.identifier.orcid0000-0002-5658-5445
person.identifier.orcid0000-0002-3096-8325
person.identifier.orcid0000-0003-1613-9675
person.identifier.orcid0000-0001-9996-2194
person.identifier.ridB-1602-2017
person.identifier.ridL-3104-2014
person.identifier.ridA-2014-2017
person.identifier.scopus-author-id36968590900
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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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