Influenza DNA vaccine purification using pHEMA cryogel support
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.6/9147 |
Resumo: | Influenza virus is a huge financial and social burden for health care systems over the world. Currently, traditionalapproaches are not effective in the fight of the epidemy and new alternatives like DNA vaccines have been developed. However, the downstream process of DNA vaccines is a constant challenge in the biotechnology industry. Cryogels has several advantages over traditional supports and have been tested as stationary phase in chromatographic separations. In this work, a method based on poly(2-hydroxyethyl methacrylate) cryogel was used to purify the plasmid NTC7482-41H-VA2 HA, which express the Influenza hemagglutinin gene. For this purpose, the cryogel was synthesized by cryo-polymerization of 2-hydroxyethyl methacrylate and characterized by scanning electron microscopy. The purification of supercoiled isoform of the plasmid NTC7482-41H-VA2 HA from a clarified lysate sample was achieved in a two-step experiment using NaCl and the dynamic binding capacity of pHEMA cryogel was determined. The assessment of DNA vaccine allowed to conclude that the level of contaminants such as proteins, genomic DNA, RNA and endotoxins are in accordance with FDA agency. |
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Influenza DNA vaccine purification using pHEMA cryogel supportInfluenzaDNA vaccinepHEMA cryogelSupercoiled purificationDNA vaccine assessmentInfluenza virus is a huge financial and social burden for health care systems over the world. Currently, traditionalapproaches are not effective in the fight of the epidemy and new alternatives like DNA vaccines have been developed. However, the downstream process of DNA vaccines is a constant challenge in the biotechnology industry. Cryogels has several advantages over traditional supports and have been tested as stationary phase in chromatographic separations. In this work, a method based on poly(2-hydroxyethyl methacrylate) cryogel was used to purify the plasmid NTC7482-41H-VA2 HA, which express the Influenza hemagglutinin gene. For this purpose, the cryogel was synthesized by cryo-polymerization of 2-hydroxyethyl methacrylate and characterized by scanning electron microscopy. The purification of supercoiled isoform of the plasmid NTC7482-41H-VA2 HA from a clarified lysate sample was achieved in a two-step experiment using NaCl and the dynamic binding capacity of pHEMA cryogel was determined. The assessment of DNA vaccine allowed to conclude that the level of contaminants such as proteins, genomic DNA, RNA and endotoxins are in accordance with FDA agency.Project POCI-01-0145-FEDER-007491ElsevieruBibliorumSantos, TiagoBrito, AndreiaBoto, RenatoSousa, PedroAlmeida, PauloCruz, CarlaTomaz, C. T.2022-06-02T00:30:15Z2018-06-022018-06-02T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.6/9147eng10.1016/j.seppur.2018.06.002info: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:RCAAP2023-12-15T09:49:55Zoai:ubibliorum.ubi.pt:10400.6/9147Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:49:21.961584Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Influenza DNA vaccine purification using pHEMA cryogel support |
title |
Influenza DNA vaccine purification using pHEMA cryogel support |
spellingShingle |
Influenza DNA vaccine purification using pHEMA cryogel support Santos, Tiago Influenza DNA vaccine pHEMA cryogel Supercoiled purification DNA vaccine assessment |
title_short |
Influenza DNA vaccine purification using pHEMA cryogel support |
title_full |
Influenza DNA vaccine purification using pHEMA cryogel support |
title_fullStr |
Influenza DNA vaccine purification using pHEMA cryogel support |
title_full_unstemmed |
Influenza DNA vaccine purification using pHEMA cryogel support |
title_sort |
Influenza DNA vaccine purification using pHEMA cryogel support |
author |
Santos, Tiago |
author_facet |
Santos, Tiago Brito, Andreia Boto, Renato Sousa, Pedro Almeida, Paulo Cruz, Carla Tomaz, C. T. |
author_role |
author |
author2 |
Brito, Andreia Boto, Renato Sousa, Pedro Almeida, Paulo Cruz, Carla Tomaz, C. T. |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
uBibliorum |
dc.contributor.author.fl_str_mv |
Santos, Tiago Brito, Andreia Boto, Renato Sousa, Pedro Almeida, Paulo Cruz, Carla Tomaz, C. T. |
dc.subject.por.fl_str_mv |
Influenza DNA vaccine pHEMA cryogel Supercoiled purification DNA vaccine assessment |
topic |
Influenza DNA vaccine pHEMA cryogel Supercoiled purification DNA vaccine assessment |
description |
Influenza virus is a huge financial and social burden for health care systems over the world. Currently, traditionalapproaches are not effective in the fight of the epidemy and new alternatives like DNA vaccines have been developed. However, the downstream process of DNA vaccines is a constant challenge in the biotechnology industry. Cryogels has several advantages over traditional supports and have been tested as stationary phase in chromatographic separations. In this work, a method based on poly(2-hydroxyethyl methacrylate) cryogel was used to purify the plasmid NTC7482-41H-VA2 HA, which express the Influenza hemagglutinin gene. For this purpose, the cryogel was synthesized by cryo-polymerization of 2-hydroxyethyl methacrylate and characterized by scanning electron microscopy. The purification of supercoiled isoform of the plasmid NTC7482-41H-VA2 HA from a clarified lysate sample was achieved in a two-step experiment using NaCl and the dynamic binding capacity of pHEMA cryogel was determined. The assessment of DNA vaccine allowed to conclude that the level of contaminants such as proteins, genomic DNA, RNA and endotoxins are in accordance with FDA agency. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-06-02 2018-06-02T00:00:00Z 2022-06-02T00:30:15Z |
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/10400.6/9147 |
url |
http://hdl.handle.net/10400.6/9147 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1016/j.seppur.2018.06.002 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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 |
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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) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799136385900216320 |