Designing new downstream processes based in FPLC and ABS methodologies

• Autor(a) principal: Santos, João Henrique Picado Madalena
• 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/10773/13852
• topic: Biotecnologia industrial; Soluções iónicas; Sistemas aquosos bifásicos; Electrólitos aquosos; Cromatografia de afinidade; Compostos bioactivos - Separação (Tecnologia)

The main objective of this work was the design of novel downstream processes for analytical and biotechnological applications. This work was divided in three major topics: formulation of novel polymer-based ABS systems with innovative use of IL as electrolytes; design of aqueous biphasic systems as an alternative for extraction of benzoylecgnonine and two hallucinogens (harmine and harmaline); FPLC purification of L-Asparaginase I. For each chapter, a detailed summary is presented. The downstream processes herein studied namely ABS and FPLC are good alternatives, due to the high yields and enhanced selectivity obtained. Moreover, they are relatively simple and fast from the operational point of view, present a low cost and are easy to scale up. The analytical studies show that further work is required for the application of ABS as drug extracting platforms, mainly because of the very low concentration of drugs in complex matrixes, such as urine and Ayahuasca. Better results were obtained using ABS as biotechnological extracting platforms, being possible to maximize the extractive performance of Cyt c and CA, using low concentrations of IL (0.1 wt%) as electrolytes. Furthermore, the FPLC purification of His-Tagg recombinant ASPase I was successfully performed, obtaining a recovery of 81.03 ± 0.01 % and a purification factor of 17 times relatively to the original sample from fermentation broth.