Candidate-based biomarker verification relies upon specific, quantitative assays optimized for selective detection of target proteins, and is increasingly viewed as a critical step in the biomarker discovery pipeline that bridges unbiased discovery to pre-clinical validation. Although individual laboratories have demonstrated that multiple reaction monitoring (MRM) coupled with isotope dilution mass spectrometry has suitable assay performance for quantitative measurements of candidate protein biomarkers in plasma, reproducibility and transferability of these assays across the biomarker community has not been demonstrated. Here we describe a multi-laboratory study to assess performance metrics of multiplexed, MRM-based assays, including reproducibility, recovery, and limits of detection and quantitation. Using common materials and standardized protocols, we demonstrate that MS-based assays of proteins in plasma can be sensitive and highly reproducible across laboratories and instrument platforms. We provide data and benchmarks against which individual laboratories can compare their performance and evaluate new technologies for biomarker verification in plasma.
Citation: Nature Biotechnology
Pub Type: Journals
proteomics, NCI, CPTAC, LC/MS/MS, quantification