Monolith and cryogel supports as new approaches to nucleic acids purification

• Autor(a) principal: Ribeiro, João Ferreira
• Data de Publicação: 2021
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10400.6/12900
• topic: Criogéis; Cromatografia; Dna; Dna Plasmídico; Dna Plasmídico Superenrolado; Monólitos; Rna; Terapia Genética; Domínio/Área Científica::Engenharia e Tecnologia::Biotecnologia

In recent years, advances in the study of the human genome and proteome have provided essential information regarding the correct functioning of genes, as well as the impact of gene mutations on human health. Gene therapy has been shown to be one of the most promising tools for the treatment of genetic or acquired diseases. Although viral vectors are currently the most used, plasmid DNA has been shown to be a strong alternative for the treatment of these diseases. To obtain plasmid DNA (pDNA) in supercoiled form, it is necessary to purify complex extracts, in which some molecules share similar characteristics and properties, which in addition can cause adverse reactions in the human body. The removal of these impurities involves costly processes and potentially harmful to the environment due to the use of organic solvents and salts. To make the purification of nucleic acids more sustainable, alternative techniques such as the use of monoliths and cryogels have been explored. Monoliths are based on organic and inorganic polymers and consist of a single piece of a highly interconnected network of channels, with mesopores and macropores, making them different from other supports. Cryogels are gel matrices prepared at freezing temperatures, characterized by a system of large and interconnected pores. Both types of supporst have advantages over conventional particulate supports, such as greater capacity and reproducibility, high flow rates and ease of scale-up. This dissertation consists of three chapters that include an introduction, a review article, and the conclusions and future perspectives. The first chapter presents a short introduction on the application of nucleic acids for the treatment of diseases, their vectors, forms of production and purification. The second chapter consists of a review article that addresses new approaches to the use of cryogels and monoliths, as an alternative to traditional chromatographic methods for the purification of nucleic acids. The third chapter presents the conclusions of this dissertation and future perspectives on the topic discussed.