title: Lentivirus capture directly from cell culture with Q-functionalised microcapillary film chromatography.
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.
authors: Department of Chemical Engineering and Biotechnology, New Museums Site, Pembroke St, Cambridge CB2 3RA, UK. Electronic address: nickdarton@cantab.net.
authors: N J
authors: NJ
authors: Darton
authors: King's College London, 123 Coldharbour Lane, London SE5 9NU, UK.
authors: D
authors: D
authors: Darling
authors: Department of Chemical Engineering and Biotechnology, New Museums Site, Pembroke St, Cambridge CB2 3RA, UK.
authors: M J
authors: MJ
authors: Townsend
authors: Department of Chemical Engineering and Biotechnology, New Museums Site, Pembroke St, Cambridge CB2 3RA, UK.
authors: D J
authors: DJ
authors: McNally
authors: King's College London, 123 Coldharbour Lane, London SE5 9NU, UK.
authors: F
authors: F
authors: Farzaneh
authors: Department of Chemical Engineering and Biotechnology, New Museums Site, Pembroke St, Cambridge CB2 3RA, UK.
authors: N K H
authors: NKH
authors: Slater
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors / Q000302:isolation & purification*; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000302:isolation & purification*; D011145:Polyvinyls / Q000737:chemistry
chemical_list: D011145:Polyvinyls; C020320:ethylene-vinyl alcohol copolymer