Purification of foamy viral particles.
Foamy viruses are non-pathogenic retroviruses and represent a tool for vector development. For gene therapy applications and for analyses of viral protein composition infectious particles need to be purified, which has been difficult for foamy viruses in the past. Here, we describe a novel, simple, and fast purification method for prototype foamy viruses with high purity using size exclusion and affinity chromatography. More than 99,9% of the contaminating proteins were removed. The purified viruses were used to determine the amount of the incorporated Pol protein relative to Gag. The determined Gag to Pol PR-RT ratio of 30:1 confirmed previous studies suggesting FV virions encapsidate fewer number of Pol molecules than orthoretroviruses.
Institut für Virologie und Immunbiologie, Julius-Maximilians-Universität Würzburg, Germany.
Ralf
R
Spannaus
Institut für Virologie und Immunbiologie, Julius-Maximilians-Universität Würzburg, Germany.
Christina
C
Miller
Institut für Virologie, Technische Universität Dresden, Germany.
Dirk
D
Lindemann
Institut für Virologie und Immunbiologie, Julius-Maximilians-Universität Würzburg, Germany. Electronic address: Jochen.Bodem@vim.uni-wuerzburg.de.
Jochen
J
Bodem
D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D015967:Gene Expression Regulation, Viral; D015684:Gene Products, pol / Q000235:genetics / Q000378:metabolism; D006801:Humans; D012192:Retroviridae Infections / Q000821:virology; D016092:Spumavirus / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification / Q000502:physiology; D014771:Virion / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification / Q000502:physiology; D019065:Virus Assembly; D014776:Virus Cultivation
D015684:Gene Products, pol
Foamy virus; Particle composition; Virus chromatography; Virus purification