Concentração de partículas pseudovirais de Zika vírus por ultrafiltração

• Ano de defesa: 2019
• Autor(a) principal: Zamprognio, Maycou Soares
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal do Rio de Janeiro; Brasil; Instituto Alberto Luiz Coimbra de Pós-Graduação e Pesquisa de Engenharia; Programa de Pós-Graduação em Engenharia Química; UFRJ
• Palavras-chave em Português: Flavivírus; Zika; Partícula pseudoviral; Concentração; CNPQ::ENGENHARIAS::ENGENHARIA QUIMICA::PROCESSOS INDUSTRIAIS DE ENGENHARIA QUIMICA::PROCESSOS BIOQUIMICOS
• Link de acesso: http://hdl.handle.net/11422/20257

This work assessed the separation of virus-like particles (VLPs) through ultrafiltration operation. The VLPs were constitutively expressed by HEK293 - 3F6 cell line (Human Embryonic Kidney cells) transfected with commercial plasmidial vector PCINEO® (Promega) encoding the genetic sequences of premembrane (pr-M) and envelope (E) structural proteins of Zika virus. The assessment techniques were based on qualitative analysis, such as the Immunoblot test, SDS-PAGE and Western blot; protein quantification, through densitometry and Bradford assays; and the particle diameter distribution analysis by using HPLC/SEC-UV. Sequential batches of 14 mL in Vivaspin® devices embedded with polyethersulfone of 300 kDa and 1000 kDa membranes were analyzed and better performance was achieved with the lowest cut-off. Therefore, batch operations in agitated Amicon® cell filled with 200 mL of clarified supernant were conducted with 500 kDa polyethersulfone and 100 kDa regenerated cellulose membranes. Despite both membranes showing concentration capacity to Zika virus-like particles, the PES membrane displayed higher efficiency.