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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.

SUBMITTER: Alqahtani A 

PROVIDER: S-EPMC4202269 | biostudies-literature | 2014 Nov

REPOSITORIES: biostudies-literature

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

Alqahtani Ali A   Heesom Kate K   Bramson Jonathan L JL   Curiel David D   Ugai Hideyo H   Matthews David A DA  

The Journal of general virology 20140805 Pt 11


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  ...[more]