Proteomics

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Adenovirus particles SILAC analysis of different batches and mutant virus


ABSTRACT: We used stable isotope labelling of amino acids in cell culture coupled with high throughput quantitative mass spectrometry to analyse the protein composition of highly purified 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 which had been used in a clinical trail. We found that the viral protein abundance and composition was 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 makes a powerful method to analyse the protein composition of this important gene therapy vector and genetically engineered or synthetic virus like particles.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

DISEASE(S): Adenovirus Infectious Disease

SUBMITTER: David Matthews  

LAB HEAD: David Matthews

PROVIDER: PXD001120 | Pride | 2014-08-08

REPOSITORIES: Pride

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Publications

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]