Production of membranes for filtration of biomolecules
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.6/4723 |
Resumo: | Plasmids are autonomously replicating entities that can be found in all bacterial species and contribute for bacterial adaption and evolution. The demand of highly purified biomolecules has triggered the development of new separation technologies. Herein, plasmid DNA (pDNA) purification process has been extensively investigated, in order to obtain highly purified molecules for gene therapy applications and DNA (deoxyribonucleic acid) vaccines. The purification of plasmid DNA is currently performed by different techniques, namely chromatography (anion-exchange, hydrophobic interaction, reversed phase, affinity and sizeexclusion), enzymatic and membrane processes. Membrane technology is a broad and highly interdisciplinary field, where process engineering, material science and chemistry meet to produce membranes that have a wide range of applications, such as water, biomolecules and plasmid DNA purification. Furthermore, membrane systems take advantage of their selectivity, high surface-area-per-unit-volume. Herein, the main goal was to produce membrane systems - electrospinning membranes: poly ε-caprolactone, polyethylene oxide and k-carrageenan; modified-nylon membranes: nylon membrane impregnated with agarose - in order to perform microfiltration and ultrafiltration processes, respectively. The produced membranes were characterized by Scanning Electron Microscopy, Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy and Energy Dispersive Spectroscopy. The water contact angles were also determined and the results obtained showed that the produced membranes presented a hydrophilic character, which is in agreement with the date previously described in literature. Porosity studies were also performed and the results demonstrated that the electrospun membranes have porosity around to 80% and the modified-nylon membranes have porosities of approximately 40%. These values can be considered to be very high, when comparing these membranes to conventional microfiltration and ultrafiltration membranes produced by phase inversion. The plasmid DNA rejection was determined on the membranes produced and the experimental results showed that the modified-nylon membrane presented rejection values up to 100%, depending on the imposed permeate flux, which is an attractive feature for its application on plasmid DNA purification by ultrafiltration. In respect to the electrospun membranes produced the observed rejections were found to be lower, up to 30%, which demonstrates that this membranes need to be optimized or modified (post-synthesis modification). |
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Production of membranes for filtration of biomoleculesBiomoleculesElectrospinningMembrane modificationMicrofiltrationPlasmid DNAPurificationUltrafiltrationDomínio/Área Científica::Engenharia e Tecnologia::Engenharia QuímicaPlasmids are autonomously replicating entities that can be found in all bacterial species and contribute for bacterial adaption and evolution. The demand of highly purified biomolecules has triggered the development of new separation technologies. Herein, plasmid DNA (pDNA) purification process has been extensively investigated, in order to obtain highly purified molecules for gene therapy applications and DNA (deoxyribonucleic acid) vaccines. The purification of plasmid DNA is currently performed by different techniques, namely chromatography (anion-exchange, hydrophobic interaction, reversed phase, affinity and sizeexclusion), enzymatic and membrane processes. Membrane technology is a broad and highly interdisciplinary field, where process engineering, material science and chemistry meet to produce membranes that have a wide range of applications, such as water, biomolecules and plasmid DNA purification. Furthermore, membrane systems take advantage of their selectivity, high surface-area-per-unit-volume. Herein, the main goal was to produce membrane systems - electrospinning membranes: poly ε-caprolactone, polyethylene oxide and k-carrageenan; modified-nylon membranes: nylon membrane impregnated with agarose - in order to perform microfiltration and ultrafiltration processes, respectively. The produced membranes were characterized by Scanning Electron Microscopy, Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy and Energy Dispersive Spectroscopy. The water contact angles were also determined and the results obtained showed that the produced membranes presented a hydrophilic character, which is in agreement with the date previously described in literature. Porosity studies were also performed and the results demonstrated that the electrospun membranes have porosity around to 80% and the modified-nylon membranes have porosities of approximately 40%. These values can be considered to be very high, when comparing these membranes to conventional microfiltration and ultrafiltration membranes produced by phase inversion. The plasmid DNA rejection was determined on the membranes produced and the experimental results showed that the modified-nylon membrane presented rejection values up to 100%, depending on the imposed permeate flux, which is an attractive feature for its application on plasmid DNA purification by ultrafiltration. In respect to the electrospun membranes produced the observed rejections were found to be lower, up to 30%, which demonstrates that this membranes need to be optimized or modified (post-synthesis modification).Os plasmídeos são entidades auto-replicantes que podem ser encontrados em todas as espécies de bactérias e que têm um papel fundamental na adaptação e evolução das bactérias. A necessidade de obter biomoléculas com um elevado grau de pureza desencadeou o desenvolvimento de novas técnicas de separação. Os processos de optimização da purificação de ADN plasmídico têm sido estudados exaustivamente, para que estes possam ser usados em aplicações de terapia génica ou em vacinas de ADN (ácido desoxirribonucleico). A purificação do ADN plasmídico tem sido realizada usando, nomeadamente a tecnologia de membranas. A tecnologia de membranas abrange uma vasta área do conhecimento, altamente interdisciplinar, onde engenharia de processos, a ciência dos materiais e a química permitem a produção de membranas aplicadas em diferentes áreas, tais como purificação de águas, biomoléculas e de ADN plasmídico, entre muitas outras. Os sistemas de separação com membranas caracterizam-se por oferecer geralmente elevada selectividade nas separações, elevadas áreas superficiais por unidade de volume do equipamento; e oferecem a possibilidade de controlar o nível de contacto e/ou mistura entre duas fases. O presente estudo teve como objectivo produzir membranas de micro e ultrafiltração para purificação de ADN plasmídico - membranas de electrospinning: poli ε-caprolactona, óxido de polietileno e k-carragenano; membranas de nylon modificadas: membrana nylon impregnada com agarose. As membranas produzidas foram caracterizadas por Microscopia Electrónica de Varrimento, Espectroscopia de Infravermelho com Transformada de Fourier Atenuada e Espectroscopia de Raio X por Dispersão de Energia. Os ângulos de contacto com água também foram determinados e os resultados obtidos mostraram que as membranas produzidas apresentam caracter hidrofílico. Estudos de porosidade foram igualmente efectuados e os resultados demonstraram que as membranas produzidas por electrospinning têm porosidades próximas de 80% e a membrana de nylon modificada tem uma porosidade próxima de 40%. Estes valores podem ser considerados bastante elevados, se os compararmos com valores típicos de membranas de microfiltração e ultrafiltração convencionais, produzidas por inversão de fase. A rejeição de ADN plasmídico foi determinada para ambas as membranas. Os valores obtidos mostraram que a membrana de nylon modificada apresentou valores de rejeição até 100%, dependendo do fluxo de permeação imposto, o que é uma característica promissora para a sua aplicação em purificação de ADN plasmídico por ultrafiltração. Relativamente às membranas produzidas por electrospinning a rejeição foi menor, apenas até 30%, o que demonstra que estas membranas precisam ser optimizadas ou eventualmente modificadas (modificação por síntese).Morão, António Miguel Parreira Cabral ForjazCorreia, Ilídio Joaquim SobreirauBibliorumCastilho, Pedro Henrique Barata2018-03-28T15:59:21Z2014-10-312014-10-022014-10-31T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.6/4723TID:201328895enginfo: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:RCAAP2023-12-15T09:42:02Zoai:ubibliorum.ubi.pt:10400.6/4723Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:45:46.235343Repositório 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 |
Production of membranes for filtration of biomolecules |
title |
Production of membranes for filtration of biomolecules |
spellingShingle |
Production of membranes for filtration of biomolecules Castilho, Pedro Henrique Barata Biomolecules Electrospinning Membrane modification Microfiltration Plasmid DNA Purification Ultrafiltration Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Química |
title_short |
Production of membranes for filtration of biomolecules |
title_full |
Production of membranes for filtration of biomolecules |
title_fullStr |
Production of membranes for filtration of biomolecules |
title_full_unstemmed |
Production of membranes for filtration of biomolecules |
title_sort |
Production of membranes for filtration of biomolecules |
author |
Castilho, Pedro Henrique Barata |
author_facet |
Castilho, Pedro Henrique Barata |
author_role |
author |
dc.contributor.none.fl_str_mv |
Morão, António Miguel Parreira Cabral Forjaz Correia, Ilídio Joaquim Sobreira uBibliorum |
dc.contributor.author.fl_str_mv |
Castilho, Pedro Henrique Barata |
dc.subject.por.fl_str_mv |
Biomolecules Electrospinning Membrane modification Microfiltration Plasmid DNA Purification Ultrafiltration Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Química |
topic |
Biomolecules Electrospinning Membrane modification Microfiltration Plasmid DNA Purification Ultrafiltration Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Química |
description |
Plasmids are autonomously replicating entities that can be found in all bacterial species and contribute for bacterial adaption and evolution. The demand of highly purified biomolecules has triggered the development of new separation technologies. Herein, plasmid DNA (pDNA) purification process has been extensively investigated, in order to obtain highly purified molecules for gene therapy applications and DNA (deoxyribonucleic acid) vaccines. The purification of plasmid DNA is currently performed by different techniques, namely chromatography (anion-exchange, hydrophobic interaction, reversed phase, affinity and sizeexclusion), enzymatic and membrane processes. Membrane technology is a broad and highly interdisciplinary field, where process engineering, material science and chemistry meet to produce membranes that have a wide range of applications, such as water, biomolecules and plasmid DNA purification. Furthermore, membrane systems take advantage of their selectivity, high surface-area-per-unit-volume. Herein, the main goal was to produce membrane systems - electrospinning membranes: poly ε-caprolactone, polyethylene oxide and k-carrageenan; modified-nylon membranes: nylon membrane impregnated with agarose - in order to perform microfiltration and ultrafiltration processes, respectively. The produced membranes were characterized by Scanning Electron Microscopy, Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy and Energy Dispersive Spectroscopy. The water contact angles were also determined and the results obtained showed that the produced membranes presented a hydrophilic character, which is in agreement with the date previously described in literature. Porosity studies were also performed and the results demonstrated that the electrospun membranes have porosity around to 80% and the modified-nylon membranes have porosities of approximately 40%. These values can be considered to be very high, when comparing these membranes to conventional microfiltration and ultrafiltration membranes produced by phase inversion. The plasmid DNA rejection was determined on the membranes produced and the experimental results showed that the modified-nylon membrane presented rejection values up to 100%, depending on the imposed permeate flux, which is an attractive feature for its application on plasmid DNA purification by ultrafiltration. In respect to the electrospun membranes produced the observed rejections were found to be lower, up to 30%, which demonstrates that this membranes need to be optimized or modified (post-synthesis modification). |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-10-31 2014-10-02 2014-10-31T00:00:00Z 2018-03-28T15:59:21Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.6/4723 TID:201328895 |
url |
http://hdl.handle.net/10400.6/4723 |
identifier_str_mv |
TID:201328895 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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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) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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