Scientific Publications

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

Publication TypeJournal Article
AuthorsAddona, TA, Abbatiello SE, Schilling B., Skates SJ, Mani D. R., Bunk DM, Spiegelman CH, Zimmerman LJ, Ham AJ, 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., Anderson NL, Fisher SJ, Liebler DC, Paulovich AG, Regnier FE, Tempst P., and Carr SA
AbstractVerification 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.
Year of Publication2009
JournalNature biotechnology
Date Published (YYYY/MM/DD)2009/07/01
ISSN Number1087-0156