Scientific Publications

Interlaboratory study characterizing a yeast performance standard for benchmarking LC-MS platform performance.

Publication TypeJournal Article
AuthorsPaulovich, AG, Billheimer D., Ham AJ, Vega-Montoto L., Rudnick PA, Tabb DL, Wang P., Blackman RK, Bunk DM, Cardasis HL, Clauser KR, Kinsinger CR, Schilling B., Tegeler TJ, Variyath AM, Wang M., Whiteaker JR, Zimmerman LJ, Fenyo D., Carr SA, Fisher SJ, Gibson BW, Mesri M., Neubert TA, Regnier FE, Rodriguez H., Spiegelman C., Stein SE, Tempst P., and Liebler DC
AbstractOptimal performance of LC-MS/MS platforms is critical to generating high quality proteomics data. Although individual laboratories have developed quality control samples, there is no widely available performance standard of biological complexity (and associated reference data sets) for benchmarking of platform performance for analysis of complex biological proteomes across different laboratories in the community. Individual preparations of the yeast Saccharomyces cerevisiae proteome have been used extensively by laboratories in the proteomics community to characterize LC-MS platform performance. The yeast proteome is uniquely attractive as a performance standard because it is the most extensively characterized complex biological proteome and the only one associated with several large scale studies estimating the abundance of all detectable proteins. In this study, we describe a standard operating protocol for large scale production of the yeast performance standard and offer aliquots to the community through the National Institute of Standards and Technology where the yeast proteome is under development as a certified reference material to meet the long term needs of the community. Using a series of metrics that characterize LC-MS performance, we provide a reference data set demonstrating typical performance of commonly used ion trap instrument platforms in expert laboratories; the results provide a basis for laboratories to benchmark their own performance, to improve upon current methods, and to evaluate new technologies. Additionally, we demonstrate how the yeast reference, spiked with human proteins, can be used to benchmark the power of proteomics platforms for detection of differentially expressed proteins at different levels of concentration in a complex matrix, thereby providing a metric to evaluate and minimize pre-analytical and analytical variation in comparative proteomics experiments.
Year of Publication2010
JournalMolecular & cellular proteomics : MCP
Volume9
Issue2
Pages242-54
Date Published (YYYY/MM/DD)2010/02/01
ISSN Number1535-9476
DOI10.1074/mcp.M900222-MCP200
PubMedhttp://www.ncbi.nlm.nih.gov/pubmed/19858499?dopt=Abstract