Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.6/2772 |
Resumo: | Nowadays, over 30 million people suffer from dementia worldwide and, with no cure in sight, these numbers tend to increase very fast. Of all dementing disorders, Alzheimer's disease (AD) is the most predominant. In AD there is a widespread synaptic and neuronal loss that causes a progressive decline in memory and other cognitive functions ultimately leading to dementia. This neurodegenerative disease is associated with two types of protein deposits, extracellular amyloid-β (Aβ) plaques and intraneuronal neurofibrillary tangles (NFTs). Recent studies demonstrated that the miR-29 cluster can decrease the production of the Aβ plaques by acting directly on the β-amyloid cleavage enzyme (BACE-1), which is critical for the cleavage of amyloid precursor protein and the generation of toxic Aβ species. These studies raised the possibility of using pre-miR-29 as a novel target for therapeutic intervention on AD, being essential to guarantee the purity, stability and integrity of the miRNA. Hence, the main goal of this study is the development of a new affinity chromatographic strategy with an Ophospho-L-tyrosine matrix, comparing two different methodologies – experimental design versus “one at a time” method - to obtain the pre-miR29 with high purity degree and yield, envisioning their application in gene therapy. |
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Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbDCromatografia de afinidadeMatrix O-phospho-L-tyrosineNowadays, over 30 million people suffer from dementia worldwide and, with no cure in sight, these numbers tend to increase very fast. Of all dementing disorders, Alzheimer's disease (AD) is the most predominant. In AD there is a widespread synaptic and neuronal loss that causes a progressive decline in memory and other cognitive functions ultimately leading to dementia. This neurodegenerative disease is associated with two types of protein deposits, extracellular amyloid-β (Aβ) plaques and intraneuronal neurofibrillary tangles (NFTs). Recent studies demonstrated that the miR-29 cluster can decrease the production of the Aβ plaques by acting directly on the β-amyloid cleavage enzyme (BACE-1), which is critical for the cleavage of amyloid precursor protein and the generation of toxic Aβ species. These studies raised the possibility of using pre-miR-29 as a novel target for therapeutic intervention on AD, being essential to guarantee the purity, stability and integrity of the miRNA. Hence, the main goal of this study is the development of a new affinity chromatographic strategy with an Ophospho-L-tyrosine matrix, comparing two different methodologies – experimental design versus “one at a time” method - to obtain the pre-miR29 with high purity degree and yield, envisioning their application in gene therapy.Atualmente, mais de 30 milhões de pessoas sofrem de demência em todo o mundo e, sem tratamento visível num futuro próximo, os números tendem a aumentar rapidamente. De todas as doenças neurodegenerativas, a doença de Alzheimer (DA) é a mais predominante. Na Doença de Alzheimer há uma perda neuronal e sináptica generalizada que causa um declínio progressivo na memória e em outras funções cognitivas, conduzindo finalmente à demência. Esta doença neurodegenerativa está associada a dois tipos de depósitos proteicos, placas extracelulares β-amilóides (Aβ) e complexos neurofibrilares intraneuronais. Estudos recentes demonstraram que o cluster miR-29 pode diminuir a deposição das placas de Aβ atuando diretamente sobre a enzima que cliva os péptidos β-amilóides (BACE-1), a qual é responsável pela clivagem da proteína precursora e, consequentemente geração de espécies Aβ tóxicas. Estes estudos apresentam a possibilidade da utilização do pré-miR-29 como um novo alvo terapêutico no tratamento da DA, para qual é necessário garantir um elevado grau de pureza, estabilidade e integridade do miRNA. Assim, o principal objetivo deste estudo é o desenvolvimento de uma nova estratégia de cromatografia de afinidade com uma matriz Ofosfo-L-tirosina comparando duas metodologias diferentes – o desenho experimental e o método “tentativa/erro” - para obter o pré-miR29 com elevado grau de pureza e rendimento, visando a sua aplicação em terapia génica.Sousa, Fani Pereira deSousa, Ângela Maria Almeida deuBibliorumAfonso, Adriana Alexandra Fernandes2014-12-09T22:29:19Z2013-0620132013-06-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.6/2772enginfo: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:RCAAP2023-12-15T09:38:56Zoai:ubibliorum.ubi.pt:10400.6/2772Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T00:44:23.398156Repositório 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 |
Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD |
title |
Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD |
spellingShingle |
Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD Afonso, Adriana Alexandra Fernandes Cromatografia de afinidade Matrix O-phospho-L-tyrosine |
title_short |
Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD |
title_full |
Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD |
title_fullStr |
Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD |
title_full_unstemmed |
Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD |
title_sort |
Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD |
author |
Afonso, Adriana Alexandra Fernandes |
author_facet |
Afonso, Adriana Alexandra Fernandes |
author_role |
author |
dc.contributor.none.fl_str_mv |
Sousa, Fani Pereira de Sousa, Ângela Maria Almeida de uBibliorum |
dc.contributor.author.fl_str_mv |
Afonso, Adriana Alexandra Fernandes |
dc.subject.por.fl_str_mv |
Cromatografia de afinidade Matrix O-phospho-L-tyrosine |
topic |
Cromatografia de afinidade Matrix O-phospho-L-tyrosine |
description |
Nowadays, over 30 million people suffer from dementia worldwide and, with no cure in sight, these numbers tend to increase very fast. Of all dementing disorders, Alzheimer's disease (AD) is the most predominant. In AD there is a widespread synaptic and neuronal loss that causes a progressive decline in memory and other cognitive functions ultimately leading to dementia. This neurodegenerative disease is associated with two types of protein deposits, extracellular amyloid-β (Aβ) plaques and intraneuronal neurofibrillary tangles (NFTs). Recent studies demonstrated that the miR-29 cluster can decrease the production of the Aβ plaques by acting directly on the β-amyloid cleavage enzyme (BACE-1), which is critical for the cleavage of amyloid precursor protein and the generation of toxic Aβ species. These studies raised the possibility of using pre-miR-29 as a novel target for therapeutic intervention on AD, being essential to guarantee the purity, stability and integrity of the miRNA. Hence, the main goal of this study is the development of a new affinity chromatographic strategy with an Ophospho-L-tyrosine matrix, comparing two different methodologies – experimental design versus “one at a time” method - to obtain the pre-miR29 with high purity degree and yield, envisioning their application in gene therapy. |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013-06 2013 2013-06-01T00:00:00Z 2014-12-09T22:29:19Z |
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/10400.6/2772 |
url |
http://hdl.handle.net/10400.6/2772 |
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.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 |
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1799136340969783296 |