Extração de adenovírus em sistemas micelares de duas fases aquosas

Abstract

Palavras-chave:Extração líquido-líquido. Adenovirus. Sistema micelar de duas fases aquosas. Biopartículas. ABSTRACTBiotechnological processes depend significantly on separation and purification techniques used to maintain cost-effective industrial-scale production of biotechnological products for commercial, industrial and therapeutic uses. The application of the aqueous two-phase micelar system (ATPMS) is proposed as an alternative for purification, since it allows the separation and analysis of biomolecules /bioparticles, often without loses of activity or their properties. This allows to perform a selective partition that enables high yields. This work aims to study the use of this methodology in the extraction and purification of adenovirus in micelle aqueous twophase formed by TritonX-114/Viral suspension. All assays were performed in 5 g systems following a full factorial design (2 3 ) with four central points. The studied factors were extraction temperature, pH of the viral suspension and concentration of the surfactant. Systems containing masses of 3g, 10g and 40g were evaluated.Extraction procedure effects over integrity and infectivity of adenovirus were also evaluated. Some of the parameters evaluated in the viral recovery process were viral potency (R Pv ) and recovery of viral specific potency (R Pvφ ). These two parameters measure the inactivation of Adenovirus by the extraction process and both showed improvement when compared with the viral suspension for some of the systems studied (i.e RPv: 341% and RPvφ 1466%). These results show that ATPMS selectively partition the infectious viral particles. According to the results of the experimental design is possible to increase, even further, these results controlling the surfactant concentration, viral suspension pH and temperature of extraction.