title: Label-Free Kinetic Studies of Hemostasis-Related Biomarkers Including D-Dimer Using Autologous Serum Transfusion.
abstract: The objective of this study was to evaluate the elimination kinetics of hemostasis-related biomarkers including the prothrombin activation fragment F1+2, thrombin-antithrombin complex (TAT), plasmin-α2-antiplasmin complex (PAP), and D-dimer in humans. Autologous serum was used as a biomarker source and infused into 15 healthy volunteers. Serum was prepared from whole blood in the presence of recombinant tissue-type plasminogen activator (final concentration 20 μg/mL) to induce plasmin generation required for PAP and D-dimer formation. Serum transfusions (50 mL/30 min) were well tolerated by all subjects. Endogenous thrombin formation was not induced by serum infusions as measured using a highly sensitive oligonucleotide-based enzyme capture assay. Median peak levels (x-fold increase over baseline) of F1+2, TAT, PAP, and D-dimer of 3.7 nmol/L (28.9), 393 ng/mL (189.6), 3,829 ng/mL (7.0), and 13.4 mg/L (34.2) were achieved at the end of serum infusions. During a 48 h lasting follow-up period all biomarkers showed elimination kinetics of a two-compartment model. Median (interquartile range) terminal half-lives were 1.9 (1.3-3.6) h for F1+2, 0.7 (0.7-2.6) h for TAT, and 10.8 (8.8-11.4) h for PAP. With 15.8 (13.1-23.1) h the D-dimer half-life was about twice as long as previously estimated from radiolabeling studies in animals and small numbers of human subjects. The serum approach presented here allows label-free and simultaneous analysis of the elimination kinetics of various hemostasis-related biomarkers. Based on these data changes in biomarker levels could more precisely used to estimate the activity level of the hemostatic system.
authors.affiliation: Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Rühl
authors.affiliation: Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Berens
authors.affiliation: Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Winterhagen
authors.affiliation: Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Müller
authors.affiliation: Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Oldenburg
authors.affiliation: Institute of Experimental Hematology and Transfusion Medicine, University Hospital Bonn, Bonn, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Pötzsch
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000097:blood*; D001804:Blood Transfusion, Autologous; D005260:Female; D005338:Fibrin Fibrinogen Degradation Products / Q000032:analysis*; D005342:Fibrinolysis / Q000187:drug effects; D006207:Half-Life; D064368:Healthy Volunteers; D006801:Humans; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D011446:Prospective Studies; D013917:Thrombin / Q000032:analysis; D010959:Tissue Plasminogen Activator / Q000494:pharmacology; D055815:Young Adult
chemical_list: D015415:Biomarkers; D005338:Fibrin Fibrinogen Degradation Products; C036309:fibrin fragment D; D013917:Thrombin; D010959:Tissue Plasminogen Activator