Quantitative Protein Mass Spectrometry to Assess Plasma Proteins Involved in Coagulation and Fibrinolysis
Half of all patients referred to tertiary centers for suspected bleeding disorders remain undiagnosed using current diagnostic assays, highlighting their limitations. Hemostasis laboratories typically rely on screening tests that measure coagulation factor levels—such as APTT, PT, and fibrinogen—but these assays are not precise enough to account for the interindividual variability in bleeding phenotypes. Different tests often yield inconsistent results, and it remains unclear which ones most accurately reflect clinical bleeding tendencies. This uncertainty increases the risk of both underdiagnosis and overdiagnosis. There is a clear need to improve existing assays and develop new diagnostic tools that more accurately identify bleeding disorders.
Quantitative protein mass spectrometry (QPMS) is an emerging technology that enables the characterization and quantification of plasma proteins. Although promising, it is complex and requires multiple optimization steps before it can be adopted in clinical settings. Recent efforts have demonstrated the feasibility of using QPMS for the relative quantification of multiple coagulation and fibrinolysis proteins in plasma samples from both healthy individuals and patients with thrombosis. However, existing QPMS assays currently cover only a subset of known coagulation and fibrinolysis proteins, and some clinically important proteins—such as von Willebrand factor (VWF) and factor VIII (FVIII)—yield suboptimal results.
The LUMC departments of Thrombosis & Hemostasis and Clinical Chemistry are now working together to enhance QPMS assays and support their clinical application. A test for antithrombin has already been developed, and the next goal is to design a comprehensive assay that quantifies all relevant coagulation and fibrinolysis-related proteins in a single run. This will involve integrating existing assays and developing new ones for proteins not yet included. The test will be analytically and clinically validated using plasma samples from healthy volunteers and patients with unexplained bleeding tendencies.
Once validated, this novel QPMS assay is intended to serve as a second-line diagnostic tool for bleeding disorders, enabling faster and more accurate diagnoses.
