title: Optimised concentration and purification of retroviruses using membrane chromatography.
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.
authors.affiliation: Department of Chemical Engineering and Biotechnology, New Museums Site, Pembroke St, Cambridge CB2 3RA, UK. Electronic address: dmcnally@cantab.net.
authors.forename: D J
authors.initials: DJ
authors.lastname: McNally
authors.affiliation: King's College London, 123 Coldharbour Lane, London SE5 9NU, UK.
authors.forename: D
authors.initials: D
authors.lastname: Darling
authors.affiliation: King's College London, 123 Coldharbour Lane, London SE5 9NU, UK.
authors.forename: F
authors.initials: F
authors.lastname: Farzaneh
authors.affiliation: Pall Europe Limited, 5 Harbourgate Business Park, Southampton Road, Portsmouth PO6 4BQ, Hampshire, UK.
authors.forename: P R
authors.initials: PR
authors.lastname: Levison
authors.affiliation: Department of Chemical Engineering and Biotechnology, New Museums Site, Pembroke St, Cambridge CB2 3RA, UK.
authors.forename: N K H
authors.initials: NK
authors.lastname: Slater
mesh_terms: D000818:Animals; D002460:Cell Line; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry / Q000302:isolation & purification*; D009052:Leukemia Virus, Murine / Q000737:chemistry / Q000302:isolation & purification; D012190:Retroviridae / Q000302:isolation & purification*; D014764:Viral Proteins / Q000032:analysis
chemical_list: D014764:Viral Proteins
keywords: Chromatography; Gene therapy; Membrane adsorption; Retrovirus; Virus