Smart macroporous structures for the purification of viral particles

Detalhes bibliográficos
Autor(a) principal: Sousa, Margarida Bucho Nunes de
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/10362/12179
Resumo: Dissertação para obtenção do Grau de Mestre em Engenharia Química e Bioquímica
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spelling Smart macroporous structures for the purification of viral particlesBiopharmaceuticalsPurificationMonolithsPorous structureVirus particlesQ ligandsDissertação para obtenção do Grau de Mestre em Engenharia Química e BioquímicaThe increasing application of viral particles in vaccination and gene-based therapies, has led to the development of alternative and improved purification processes. Traditional purification methods include chromatographic techniques, however the chromatographic matrices used present limitations specially when aimed at the purification of large molecules. This work presents the preparation of chitosan-based monoliths using clean processes and easy functionalization techniques intending to improve Adenovirus serotype 5 (Ad5) purification. Monoliths were prepared by blending chitosan (CHT) with glycidylmethacrylate (GMA) or poly(vinyl alcohol) (PVA), using two preparation techniques, freeze-drying and a scCO2 – assisted drying process, and were subsequently functionalized with Q ligands by three different methods. In addition, monoliths blended with magnetic nanoparticles were also prepared using the same strategies to confer them a controlled magnetic response. The monoliths produced were characterized in terms of ligand immobilization yield, and evaluated for Ad5 purification. Two types of monoliths showed potential: the CHT/PVA(50:50) prepared by freeze drying and functionalized by the alternative plasma technique (M2) and the CHT/PVA(50:50) 7% monolith prepared by scCO2 – assisted drying process and functionalized by the epoxyactivation technique (M1). The amount of ligand Q immobilized on the supports was monitored by titration assays, among which the CHT/PVA(50:50) 7% M2 prepared by scCO2 – assisted drying process exhibited the highest immobilization yield (91%). Among the results for Ad5 purification, the CHT/PVA(50:50)M2 and the CHT/PVA(50:50)7% M1 resulted in a 40% and 14% of the viral particles, respectively. Protein-binding assays were conducted using bovine serum albumin (BSA) and lysozyme, to evaluate the anionic-exchange capacity of the supports. The results make us believe in the potential of the produced monoliths to be applied in chromatographic techniques. However further improvements are necessary to enhance virus binding and recovery, to obtain an improved purification process.project PTDC/EBB-BIO/118317/2010Faculdade de Ciências e TecnologiaRoque, AnaRicardo, AnaRUNSousa, Margarida Bucho Nunes de2014-06-16T14:54:39Z20142014-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/12179enginfo: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:RCAAP2024-03-11T03:47:03Zoai:run.unl.pt:10362/12179Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:20:47.398262Repositório 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 Smart macroporous structures for the purification of viral particles
title Smart macroporous structures for the purification of viral particles
spellingShingle Smart macroporous structures for the purification of viral particles
Sousa, Margarida Bucho Nunes de
Biopharmaceuticals
Purification
Monoliths
Porous structure
Virus particles
Q ligands
title_short Smart macroporous structures for the purification of viral particles
title_full Smart macroporous structures for the purification of viral particles
title_fullStr Smart macroporous structures for the purification of viral particles
title_full_unstemmed Smart macroporous structures for the purification of viral particles
title_sort Smart macroporous structures for the purification of viral particles
author Sousa, Margarida Bucho Nunes de
author_facet Sousa, Margarida Bucho Nunes de
author_role author
dc.contributor.none.fl_str_mv Roque, Ana
Ricardo, Ana
RUN
dc.contributor.author.fl_str_mv Sousa, Margarida Bucho Nunes de
dc.subject.por.fl_str_mv Biopharmaceuticals
Purification
Monoliths
Porous structure
Virus particles
Q ligands
topic Biopharmaceuticals
Purification
Monoliths
Porous structure
Virus particles
Q ligands
description Dissertação para obtenção do Grau de Mestre em Engenharia Química e Bioquímica
publishDate 2014
dc.date.none.fl_str_mv 2014-06-16T14:54:39Z
2014
2014-01-01T00:00:00Z
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/10362/12179
url http://hdl.handle.net/10362/12179
dc.language.iso.fl_str_mv eng
language eng
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 Faculdade de Ciências e Tecnologia
publisher.none.fl_str_mv Faculdade de Ciências e Tecnologia
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)
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
repository.mail.fl_str_mv
_version_ 1799137849128255488

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