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Blood Adv. 2017 Jun 20;1(15):1080-1087. doi: 10.1182/bloodadvances.2017007955. eCollection 2017 Jun 27.

Multiplexed targeted proteomic assay to assess coagulation factor concentrations and thrombosis-associated cancer.

Author information

Genome British Columbia Proteomics Centre, University of Victoria, Victoria, BC, Canada.
Center for Proteomics and Metabolomics and.
Einthoven Laboratory for Experimental Vascular Medicine, Division of Thrombosis and Hemostasis, Department of Internal Medicine, Leiden University Medical Center, Leiden, The Netherlands.
Molecular Pathology, Jewish General Hospital Proteomics Centre, Lady Davis Institute, McGill University, Montreal, QC, Canada.
Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada; and.
Department of Clinical Epidemiology, Leiden University Medical Center, Leiden, The Netherlands.


The plasma levels of pro- and anticoagulant proteins are important markers for venous thrombosis (VT) risk and can be affected by both genetic and acquired factors, including cancer. Generally, these markers are measured using activity- or antibody-based assays. Targeted proteomics with stable-isotope-labeled internal standards has proven adept at the rapid, multiplex, and precise quantification of proteins in complex biological samples such as plasma. We used liquid chromatography coupled to multiple reaction monitoring (MRM) mass spectrometry to evaluate the concentrations of 31 coagulation- and fibrinolysis-related proteins in plasma from 25 healthy controls, 25 patients with VT, and 25 patients with VT who were also diagnosed with cancer. The concentration level of 1 to 3 proteotypic peptides per protein was determined, and all samples were previously characterized using traditional antibody- or activity-based methods. When comparing the conventional and the MRM strategies, the mean Pearson correlation for the 13 proteins (covered by 36 target peptides) shared between the 2 approaches was 0.77, indicating a good correlation. Additionally, MRM offers higher sensitivity (mean regression slope, 0.81), higher multiplicity in a single run, and good ability to leverage all measurements to discriminate groups using unsupervised clustering, which identified vitamin K antagonist users as well as patients with VT and cancer. The data collected using MRM show that the combination of coagulation factor levels yields signature information on VT and cancer, which was not obvious from a single measurement. These results encourage the further validation and investigation of MRM in profiling protein signature of disease.

Conflict of interest statement

Conflict-of-interest disclosure: C.H.B. is the chief strategy officer of MRM Proteomics, Inc. The remaining authors declare no competing financial interests.

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