Influenza DNA vaccine purification using pHEMA cryogel support

Detalhes bibliográficos
Autor(a) principal: Santos, Tiago
Data de Publicação: 2018
Outros Autores: Brito, Andreia, Boto, Renato, Sousa, Pedro, Almeida, Paulo, Cruz, Carla, Tomaz, C. T.
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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spelling 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
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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
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dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
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