title: Analysis of purified wild type and mutant adenovirus particles by SILAC based quantitative proteomics.
abstract: We used SILAC (stable isotope labelling of amino acids in cell culture) and high-throughput quantitative MS mass spectrometry to analyse the protein composition of highly purified WT wild type adenoviruses, mutant adenoviruses lacking an internal protein component (protein V) and recombinant adenoviruses of the type commonly used in gene therapy, including one virus that had been used in a clinical trial. We found that the viral protein abundance and composition were consistent across all types of virus examined except for the virus lacking protein V, which also had reduced amounts of another viral core protein, protein VII. In all the samples analysed we found no evidence of consistent packaging or contamination with cellular proteins. We believe this technique is a powerful method to analyse the protein composition of this important gene therapy vector and genetically engineered or synthetic virus-like particles. The raw data have been deposited at proteomexchange, identifer PXD001120.
authors.affiliation: College of Applied Medical Sciences, Najran University, Najran 1983, Saudi Arabia.|School of Cellular and Molecular Medicine, University Walk, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: Ali
authors.initials: A
authors.lastname: Alqahtani
authors.affiliation: Proteomics Facility, Faculty of Medical and Veterinary Sciences, University Walk, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: Kate
authors.initials: K
authors.lastname: Heesom
authors.affiliation: McMaster Immunology Research Centre, 4016 Michael DeGroote Centre for Learning & Discovery, McMaster University, Hamilton, L8S 4L8 Ontario, Canada.
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Bramson
authors.affiliation: Biologic Therapeutics Center, School of Medicine, Washington University in St Louis, 4511 Forest Park Medical Building, St Louis, MO 63108, USA.|Cancer Biology Division, Department of Radiation Oncology, School of Medicine, Washington University in St Louis, 4511 Forest Park Medical Building, St Louis, MO 63108, USA.
authors.forename: David
authors.initials: D
authors.lastname: Curiel
authors.affiliation: Cancer Biology Division, Department of Radiation Oncology, School of Medicine, Washington University in St Louis, 4511 Forest Park Medical Building, St Louis, MO 63108, USA.
authors.forename: Hideyo
authors.initials: H
authors.lastname: Ugai
authors.affiliation: School of Cellular and Molecular Medicine, University Walk, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: David A
authors.initials: DA
authors.lastname: Matthews
mesh_terms: D000260:Adenoviruses, Human / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D000596:Amino Acids / Q000032:analysis; D002460:Cell Line; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D007553:Isotope Labeling; D009154:Mutation; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D000596:Amino Acids; D014764:Viral Proteins
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