학술논문
Multi-site assessment of the precision and reproducibility of multiple reaction monitoring–based measurements of proteins in plasma.
Document Type
Article
Author
Addona, Terri A.; Abbatiello, Susan E.; Schilling, Birgit; Skates, Steven J.; Mani, D. R.; Bunk, David M.; Spiegelman, Clifford H.; Zimmerman, Lisa J.; Ham, Amy-Joan L.; Keshishian, Hasmik; Hall, Steven C.; Allen, Simon; Blackman, Ronald K.; Borchers, Christoph H.; Buck, Charles; Cardasis, Helene L.; Cusack, Michael P.; Dodder, Nathan G.; Gibson, Bradford W.; Held, Jason M.
Source
Subject
*BIOMARKERS
*MASS spectrometry
*PROTEINS
*THERAPEUTICS
*BODY fluids
*CELLS
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Language
ISSN
1087-0156
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 μg/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. [ABSTRACT FROM AUTHOR]