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Goals: The sPRG 2009 focused on evaluating the ability of proteomics laboratories to determine the quantities of several proteins in a digest. Participation in this study aimed at providing to each laboratory with an opportunity to assess its capabilities with regard to:
• Methodologies for protein quantification by mass spectrometry
• Bioinformatics for data analysis
An additional goal of this study was to evaluate this protein mixture as the first step in the development of a quantitative analysis protein standard that can be used by the proteomics community. Each laboratory requesting a study sample received a mixture of 3–6 proteins that have been digested with Trypsin and a mixture of known quantities of 3–5 stable isotope labeled, synthetic peptide analogs corresponding to each protein. Participants were asked to determine the quantity of each protein present in the mixture based on comparison to the corresponding peptides.
Composition of the sample
- 5 purified recombinant proteins in specified amounts were digested with trypsin (unlabeled or natural peptides) and
- Spiked with selected stable isotope labeled peptide analogs (synthetic or heavy peptides).
Figure 1: ProteoRed members participating in the spRG2009 study
Figure 3: Summary of the results obtained by three of the ProteoRed participants using different experimental approaches.
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