Development and optimization of a microRNA purification technology by affinity chromatography, using the concept of QbD

• Autor(a) principal: Afonso, Adriana Alexandra Fernandes
• 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
• topic: Cromatografia de afinidade; Matrix O-phospho-L-tyrosine

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.