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

• Published in: Journal of Chromatography A 2012-08, Vol.1251, p.236-239
• Main Authors: Darton, N.J., Darling, D., Townsend, M.J., McNally, D.J., Farzaneh, F., Slater, N.K.H.
• Format: Article
• Language: English
• Subjects: adsorbents; alcohols; anion exchange; bioreactors; Capture; cell culture; chemical structure; Chromatography; composite polymers; culture media; extrusion; Gene therapy; harvesting; Lentivirus; melting; Microcapillary; particulates; Virus
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2012.06.072

▸ New disposable adsorbent material developed for fast anion-exchange capture of nano-complexes. ▸ No sample prefiltering, clarification or pre-processing required. ▸ Consists of melt extruded plastic films containing 19 parallel microcapillaries. ▸ Capture of 1×107 infectious units of lentivirus (∼clinical trial does) from un-processed sample. ▸ Might be used to harvest lentiviral particles directly from bioreactors in clinical production. 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±10μm, 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 2.5×106infectious 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 1×107ifu 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.