Multi-site assessment of the precision and reproducibility of multiple reaction monitoring-based measurements of proteins in plasma Journal Article
| Authors: | Addona, T. A.; Abbatiello, S. E.; Schilling, B.; Skates, S. J.; Mani, D. R.; Bunk, D. M.; Spiegelman, C. H.; Zimmerman, L. J.; Ham, A. J. L.; Keshishian, H.; Hall, S. C.; Allen, S.; Blackman, R. K.; Borchers, C. H.; Buck, C.; Cardasis, H. L.; Cusack, M. P.; Dodder, N. G.; Gibson, B. W.; Held, J. M.; Hiltke, T.; Jackson, A.; Johansen, E. B.; Kinsinger, C. R.; Li, J.; Mesri, M.; Neubert, T. A.; Niles, R. K.; Pulsipher, T. C.; Ransohoff, D.; Rodriguez, H.; Rudnick, P. A.; Smith, D.; Tabb, D. L.; Tegeler, T. J.; Variyath, A. M.; Vega-Montoto, L. J.; Wahlander, Å; Waldemarson, S.; Wang, M.; Whiteaker, J. R.; Zhao, L.; Anderson, N. L.; Fisher, S. J.; Liebler, D. C.; Paulovich, A. G.; Regnier, F. E.; Tempst, P.; Carr, S. A. |
| Article Title: | Multi-site assessment of the precision and reproducibility of multiple reaction monitoring-based measurements of proteins in plasma |
| Abstract: | Verification of candidate biomarkers relies upon specific, quantitative assays optimized for selective detection of target proteins, and is increasingly viewed as a critical step in the discovery pipeline that bridges unbiased biomarker discovery to preclinical validation. Although individual laboratories have demonstrated that multiple reaction monitoring (MRM) coupled with isotope dilution mass spectrometry can quantify candidate protein biomarkers in plasma, reproducibility and transferability of these assays between laboratories have not been demonstrated. We describe a multilaboratory study to assess reproducibility, recovery, linear dynamic range and limits of detection and quantification of multiplexed, MRM-based assays, conducted by NCI-CPTAC. Using common materials and standardized protocols, we demonstrate that these assays can be highly reproducible within and across laboratories and instrument platforms, and are sensitive to low g/ml protein concentrations in unfractionated plasma. We provide data and benchmarks against which individual laboratories can compare their performance and evaluate new technologies for biomarker verification in plasma. © 2009 Nature America, Inc. All rights reserved. |
| Keywords: | nonhuman; sensitivity and specificity; bio-marker discovery; common materials; critical steps; instrument platforms; isotope dilution mass spectrometry; limits of detection; linear dynamic ranges; multi-site; multiple reaction monitoring; new technologies; protein concentrations; quantitative assay; reproducibility; selective detection; target proteins; isotopes; laboratories; mass spectrometry; monitoring; plasmas; proteins; biomarkers; aprotinin; biological marker; c reactive protein; leptin; myelin basic protein; myoglobin; peroxidase; prostate specific antigen; accuracy; protein analysis; protein blood level; quality control; biological markers; blood chemical analysis; blood proteins; linear models; proteome; reproducibility of results; technology assessment, biomedical |
| Journal Title: | Nature Biotechnology |
| Volume: | 27 |
| Issue: | 7 |
| ISSN: | 1087-0156 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2009-07-01 |
| Start Page: | 633 |
| End Page: | 641 |
| Language: | English |
| DOI: | 10.1038/nbt.1546 |
| PUBMED: | 19561596 |
| PROVIDER: | scopus |
| PMCID: | PMC2855883 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 64" - "Export Date: 30 November 2010" - "CODEN: NABIF" - "Source: Scopus" |
