2009
DOI: 10.1038/nbt.1546
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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 assa… Show more

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Cited by 960 publications
(842 citation statements)
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“…SRM mass spectrometry in combination with stable isotope dilution is a powerful approach for protein quantification and offers major advantages over antibody‐based strategies (Addona et al , 2009). Not only does the method circumvent the need for antibody production, but it also provides information about absolute rather than relative amounts of protein.…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…SRM mass spectrometry in combination with stable isotope dilution is a powerful approach for protein quantification and offers major advantages over antibody‐based strategies (Addona et al , 2009). Not only does the method circumvent the need for antibody production, but it also provides information about absolute rather than relative amounts of protein.…”
Section: Discussionmentioning
confidence: 99%
“…Comprehensive studies on the proteomes of cells, tissues, or organisms have been reported (e.g. Addona et al , 2009; Nilsson et al , 2010; Beck et al , 2011; Nagaraj et al , 2011; for review, see Bensimon et al , 2012; Kulak et al , 2014; Wilhelm et al , 2014; Hein et al , 2015; Richards et al , 2015), but accurate quantitative information on proteins expressed at low levels remains scarce. As a consequence, published estimates for the abundance of specific proteins sometimes vary over several orders of magnitude.…”
Section: Introductionmentioning
confidence: 99%
“…Numerous approaches can be taken to characterize protein state in biological samples which either employ targeted mass spectrometry (MS) or indirect analysis using antibody-based techniques. MS methods such as selected reaction monitoring (SRM) 28 , also known as multiple reaction monitoring (MRM), are highly sensitive capable of quantitatively detecting attomolar concentrations of protein in complex biological samples (for review see Hjelle 29 ). Antibodybased techniques, which can detect down to femtomolar protein concentrations, include western blot, ELISA, reverse-phase protein arrays (RPPA) and flow cytometry as well as the cIEF assay described here.…”
Section: Comparison With Other Methodsmentioning
confidence: 99%
“…This program was renewed in 2011 as the CPTAC, which began focusing on applications 77. CPTAC member laboratories applied standardized methods of multiple reaction monitoring (MRM) and demonstrated reproducibility, precision, sensitive quantitation in tissues and biofluids 78. Cox and coworkers performed a similar interlaboratory precision study of IGF‐1 in plasma across 130 healthy human samples and 22 samples from patients with acromegaly, finding excellent reproducibility 79.…”
Section: Clinical Applicationsmentioning
confidence: 99%