Lentivirus capture directly from cell culture with Q-functionalised microcapillary film chromatography

J Chromatogr A. 2012 Aug 17:1251:236-239. doi: 10.1016/j.chroma.2012.06.072. Epub 2012 Jun 27.

Abstract

A new disposable adsorbent material for fast anion-exchange capture of nano-complexes without prefiltering, clarification or pre-processing of samples was developed based on plastic microcapillary films (MCFs). An MCF containing 19 parallel microcapillaries, each with a mean internal diameter of 142 ± 10 μm, was prepared using a melt extrusion process from an ethylene-vinyl alcohol copolymer (EVOH). The MCF internal surfaces were functionalised using branched chain chemistries to attach quaternary amine groups producing an anion-exchange adsorbent. The purification of nano-complexes using this newly fabricated MCF-EVOH-Q was successfully demonstrated with the capture of lentivirus from pre-filtered culture harvest. This 5m chromatographic substrate was found to bind and elute ∼40% of bound lentivirus or 2.5 × 10(6)infectious units (ifu). The unique properties of this chromatographic substrate that allow the passage of large particulates was further demonstrated with the capture of lentiviral particles from unfiltered un-processed culture media containing cells and cell debris. Using this approach, 56% or 1 × 10(7)ifu of captured lentivirus was eluted. A device based on this new material might be used at an early stage in clinical lentiviral production to harvest lentiviral particles, directly from bioreactors.

MeSH terms

  • Cell Culture Techniques / methods*
  • Cell Line, Tumor
  • Chromatography, Ion Exchange / instrumentation*
  • Chromatography, Ion Exchange / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Genetic Vectors / isolation & purification*
  • HEK293 Cells
  • Humans
  • Lentivirus / isolation & purification*
  • Polyvinyls / chemistry

Substances

  • Polyvinyls
  • ethylene-vinyl alcohol copolymer