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Nat Biotechnol DOI:10.1038/nbt.1546

Multi-site assessment of the precision and reproducibility of multiple reaction monitoring-based measurements of proteins in plasma.

Publication TypeJournal Article
Year of Publication2009
AuthorsAddona, TA, Abbatiello, SE, Schilling, B, Skates, SJ, Mani, DR, Bunk, DM, Spiegelman, CH, Zimmerman, LJ, Ham, A-JL, Keshishian, H, Hall, SC, Allen, S, Blackman, RK, Borchers, CH, Buck, C, Cardasis, HL, Cusack, MP, Dodder, NG, Gibson, BW, Held, JM, Hiltke, T, Jackson, A, Johansen, EB, Kinsinger, CR, Li, J, Mesri, M, Neubert, TA, Niles, RK, Pulsipher, TC, Ransohoff, D, Rodriguez, H, Rudnick, PA, Smith, D, Tabb, DL, Tegeler, TJ, Variyath, AM, Vega-Montoto, LJ, Wahlander, A, Waldemarson, S, Wang, M, Whiteaker, JR, Zhao, L, N Anderson, L, Fisher, SJ, Liebler, DC, Paulovich, AG, Regnier, FE, Tempst, P, Carr, SA
JournalNat Biotechnol
Date Published2009 Jul
KeywordsBiomarkers, Blood Chemical Analysis, Blood Proteins, Humans, Linear Models, Mass Spectrometry, Proteome, Reproducibility of Results, Sensitivity and Specificity, Technology Assessment, Biomedical

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 mug/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.


Alternate JournalNat. Biotechnol.
PubMed ID19561596
PubMed Central IDPMC2855883
Grant ListU24 CA126485 / CA / NCI NIH HHS / United States
U24 CA126480-01 / CA / NCI NIH HHS / United States
U24 CA126480 / CA / NCI NIH HHS / United States
U24 126479 / / PHS HHS / United States
U24 126480 / / PHS HHS / United States
U24 CA126476-03 / CA / NCI NIH HHS / United States
U24 CA126476 / CA / NCI NIH HHS / United States
U24 126477 / / PHS HHS / United States