Screening polymeric ionic liquids for chromatography-based purification of bacteriophage M13

M. J. Jacinto, Alexandra Wagner, Inês M. Sá, David J.S. Patinha, Isabel M. Marrucho, João Gonçalves, Richard C. Willson, A. M. Azevedo, M. R. Aires-Barros

Research output: Contribution to journalArticle

Abstract

M13 bacteriophage is a key instrument in phage display applications, as well as a possible antibacterial therapeutic agent due to its highly restrictive bacterial pathogenesis, and other applications. The traditional phage purification process is usually achieved by gradient ultracentrifugation or a combination of precipitation, centrifugation and microfiltration. These approaches easily lead to long process times, high operational costs, phage aggregation and consequent product loss (approximately 60%). This work is thus focused on an alternative potential large-scale process to achieve high yield and purity while minimizing the operational costs. Electrostatic-based separation processes are also common biomolecules purification techniques. Although anion exchange chromatography has been used before to purify several viral particles, this technique has been poorly reported for the purification of M13 phage. In a recent work, our group has demonstrated the use of a predominant anion exchange process, where a polymeric ionic liquid (PIL) was used as an alternative separation matrix for M13 bacteriophage. In this work, a variety of system parameters was studied, including chemical structure of the cation and the anion, the crosslinker nature and its concentration, either in batch adsorption/elution or chromatographic operation mode. The PIL-based chromatographic operation mode revealed to be a suitable separation process for M13 from directly filtered E. coli supernatant, reaching over 70% M13 recovery and 4.6 purification factor in a single step. To our knowledge, this is the first time that PILs have been reported as separation agents for bioproducts from complex mixtures.

Original languageEnglish (US)
Article number117906
JournalSeparation and Purification Technology
Volume257
DOIs
StatePublished - Feb 15 2021

Keywords

  • Anion-exchange
  • Bacteriophage M13
  • Batch adsorption
  • Chromatography
  • Polymeric ionic liquids

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

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