Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support

Detalhes bibliográficos
Autor(a) principal: Valente, J.F.A.
Data de Publicação: 2020
Outros Autores: Sousa, A., Azevedo, G.A., Queiroz, J.A., Sousa, F.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.8/8385
Resumo: This work was supported by FEDER funds through the POCI - COMPETE 2020 - Operational Programme Competitive- ness and Internationalization in Axis I - Strengthening research, technological development and innovation (Project POCI-01-0145- FEDER-007491) and National Funds by FCT - Foundation for Sci- ence and Technology (Project UID/Multi/00709/2019 ). J.F.A. Valente and A. Sousa also acknowledge Ph.D. and Postdoctoral fellowships (Ref SFRH/BD/96809/2013 and Ref SFRH/BPD/102716/2014, respec- tively).
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spelling Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous supportAffinity chromatographyArginineBiorecognitionMacroporous beadsp53-encoding plasmidSupercoiled isoformThis work was supported by FEDER funds through the POCI - COMPETE 2020 - Operational Programme Competitive- ness and Internationalization in Axis I - Strengthening research, technological development and innovation (Project POCI-01-0145- FEDER-007491) and National Funds by FCT - Foundation for Sci- ence and Technology (Project UID/Multi/00709/2019 ). J.F.A. Valente and A. Sousa also acknowledge Ph.D. and Postdoctoral fellowships (Ref SFRH/BD/96809/2013 and Ref SFRH/BPD/102716/2014, respec- tively).p53 is a tumour suppressor gene that has been explored for cancer gene therapy as a possible alter- native to the common treatments. The use of plasmid DNA (pDNA) to carry the therapeutic gene has been considered, but it is requisite to preserve its supercoiled (sc) structure, for eliciting a more effective gene expression and therapeutic action. The purification of the sc pDNA using amino acids-based affinity chromatography has been successfully applied, exploring different amino acids and supports. From these studies, it stood out the selectivity of arginine for the recognition of sc pDNA. However, some limitation on the binding capacity was found in the arginine-agarose support, and in the case of monoliths, some fouling and clogging can limit sequential runs. By using macroporous support modified with arginine it was expected to take advantage of the selectivity of the ligand combined with the flow properties and binding capacity offered by the support. The arginine-modified macroporous support was characterized by SEM, EDX and FTIR also to verify the correct immobilization of arginine, and then used for pDNA pu- rification. The support showed to be effective on the sc p53-pDNA isolation, and the robustness was also achieved by accomplishing the purification of plasmids with different sizes, only by slightly adjusting the experimental conditions. Regarding the dynamic binding capacity of the arginine-modified macrop- orous support, it was achieved an improvement of more than 50% in the pDNA binding capacity when compared with their homologous arginine-agarose commercial matrix, suggesting potential economic fea- sibility in case of scale-up.ElsevierIC-OnlineValente, J.F.A.Sousa, A.Azevedo, G.A.Queiroz, J.A.Sousa, F.2023-04-14T13:09:57Z20202020-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.8/8385engJ.F.A. Valente, A. Sousa, G.A. Azevedo, J.A. Queiroz, F. Sousa, Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support, Journal of Chromatography A, Volume 1618, 2020, 460890, ISSN 0021-9673, https://doi.org/10.1016/j.chroma.2020.4608900021-967310.1016/j.chroma.2020.460890metadata only accessinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-05-23T14:53:10ZPortal AgregadorONG
dc.title.none.fl_str_mv Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support
title Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support
spellingShingle Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support
Valente, J.F.A.
Affinity chromatography
Arginine
Biorecognition
Macroporous beads
p53-encoding plasmid
Supercoiled isoform
title_short Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support
title_full Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support
title_fullStr Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support
title_full_unstemmed Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support
title_sort Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support
author Valente, J.F.A.
author_facet Valente, J.F.A.
Sousa, A.
Azevedo, G.A.
Queiroz, J.A.
Sousa, F.
author_role author
author2 Sousa, A.
Azevedo, G.A.
Queiroz, J.A.
Sousa, F.
author2_role author
author
author
author
dc.contributor.none.fl_str_mv IC-Online
dc.contributor.author.fl_str_mv Valente, J.F.A.
Sousa, A.
Azevedo, G.A.
Queiroz, J.A.
Sousa, F.
dc.subject.por.fl_str_mv Affinity chromatography
Arginine
Biorecognition
Macroporous beads
p53-encoding plasmid
Supercoiled isoform
topic Affinity chromatography
Arginine
Biorecognition
Macroporous beads
p53-encoding plasmid
Supercoiled isoform
description This work was supported by FEDER funds through the POCI - COMPETE 2020 - Operational Programme Competitive- ness and Internationalization in Axis I - Strengthening research, technological development and innovation (Project POCI-01-0145- FEDER-007491) and National Funds by FCT - Foundation for Sci- ence and Technology (Project UID/Multi/00709/2019 ). J.F.A. Valente and A. Sousa also acknowledge Ph.D. and Postdoctoral fellowships (Ref SFRH/BD/96809/2013 and Ref SFRH/BPD/102716/2014, respec- tively).
publishDate 2020
dc.date.none.fl_str_mv 2020
2020-01-01T00:00:00Z
2023-04-14T13:09:57Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10400.8/8385
url http://hdl.handle.net/10400.8/8385
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv J.F.A. Valente, A. Sousa, G.A. Azevedo, J.A. Queiroz, F. Sousa, Purification of supercoiled p53-encoding plasmid using an arginine-modified macroporous support, Journal of Chromatography A, Volume 1618, 2020, 460890, ISSN 0021-9673, https://doi.org/10.1016/j.chroma.2020.460890
0021-9673
10.1016/j.chroma.2020.460890
dc.rights.driver.fl_str_mv metadata only access
info:eu-repo/semantics/openAccess
rights_invalid_str_mv metadata only access
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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