resumo
M13 is a filamentous, non-lytic bacteriophage that infects Escherichia coli via the F pilus. Currently, phage M13 is widely used in phage display technology and bio-nanotechnology, and is considered a possible antibacterial therapeutic agent, among other applications. Conventional phage purification involves 5-7 operational steps, with high operational costs and significant product loss (approximately 60%). In this work, we propose a scalable purification process for M13 bacteriophage using a novel stationary phase based on a polymeric ionic liquid (PIL) with a positively charged backbone structure. Poly (1-vinyl-3-ethyl imidazolium bis(trifluoromethylsulfonyl) imide) - poly(VEIM-TFSI) predominantly acted as an anion exchanger under binding-elution mode. This revealed to be a rapid and simple method for the recovery of phage M13 with an overall separation yield of over 70% after a single downstream step. To the best of our knowledge, PILs have never been used as separation matrices for biological products and the results obtained, together with the large number of cations and anions available to prepare PILs, illustrate well the large potential of the proposed methodology. (C) 2017 Elsevier B.V. All rights reserved.
palavras-chave
FILAMENTOUS BACTERIOPHAGES; IONIC LIQUIDS; PHAGE; NANOPARTICLES
categoria
Biochemistry & Molecular Biology; Chemistry
autores
Jacinto, MJ; Patinha, DJS; Marrucho, IM; Goncalves, J; Willson, RC; Azevedo, AM; Aires-Barros, MR
nossos autores
David José Souto Patinha
Estudante de doutoramento
Isabel Maria Delgado Jana Marrucho Ferreira
Professor CatedráticoGrupos
agradecimentos
This work was supported by Fundacao para Ciencia e Tecnologia (FCT, www.fct.pt) through the PhD fellowships to ME (PD/BD/105883/2014) and DJSP (SFRH/BD/97042/2013); research contract under the program "Investigador FCT 2014" to AMA (IF/00048/2014/CP1214/CT0010), research project PureMAb (PTDC/QEQ-PRS/0286/2014) and units research contracts to IBB (UID/BIO/04565/2013) and Centro de Quimica Estrutural (UID/QUI/00100/2013).