Optimised concentration and purification of retroviruses using membrane chromatography

J Chromatogr A. 2014 May 2;1340(100):24-32. doi: 10.1016/j.chroma.2014.03.023. Epub 2014 Mar 15.

Abstract

The ability of an anion exchange membrane to purify a γ-retrovirus was assessed and optimised with respect to different loading and wash buffers. Recoveries of infectious virus greater than 50% were consistently obtained, while specific titre was increased up to one thousand fold when compared to the material loaded. Specific proteins removed and retained by this optimised process were identified by mass spectrometry. It was possible to successfully bind and elute the equivalent of 1.27 × 10(8) Ifu/ml of ion exchange membrane. This could then be highly concentrated, with infectious virus concentrated to a maximum of 420-fold compared to the load.

Keywords: Chromatography; Gene therapy; Membrane adsorption; Retrovirus; Virus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Chromatography, Ion Exchange / methods*
  • Genetic Vectors / chemistry
  • Genetic Vectors / isolation & purification*
  • Leukemia Virus, Murine / chemistry
  • Leukemia Virus, Murine / isolation & purification
  • Retroviridae / isolation & purification*
  • Viral Proteins / analysis

Substances

  • Viral Proteins