Analysis of purified wild type and mutant adenovirus particles by SILAC based quantitative proteomics

Ali Alqahtani, Kate Heesom, Jonathan L Bramson, David Curiel, Hideyo Ugai, David A Matthews

Research output: Contribution to journalArticle (Academic Journal)peer-review

9 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)2504-11
Number of pages8
JournalJournal of General Virology
Volume95
Issue numberPt 11
DOIs
Publication statusPublished - Nov 2014

Bibliographical note

© 2014 The Authors.

Keywords

  • Adenoviruses, Human
  • Amino Acids
  • Cell Line
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Isotope Labeling
  • Mutation
  • Proteomics
  • Tandem Mass Spectrometry
  • Viral Proteins

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