Contact activation of the plasma coagulation cascade. III. Biophysical aspects of thrombin-binding anticoagulants.
A biophysical model linking fibrin polymerization kinetics (following release from a thrombin-fibrinogen complex), coagulation time, and competitive inhibition of thrombin illustrates the utility of thrombin-binding ligands as anticoagulants in blood collection applications. The resulting mathematical relationship connecting fibrinogen, ligand, and thrombin concentrations was tested against experimentally observed anticoagulation of whole, platelet-poor porcine plasma induced by short, single-stranded DNA oligonucleotides originally found to bind thrombin by screening combinatorial libraries. The thrombin-fibrinogen dissociation constant Ks served as the single adjustable parameter in a least-squares fitting of the model to experimental anticoagulation data. Best-fit Ks values corroborated microM values measured in plasma-free systems, and application of the model to a ligand challenge to the intrinsic pathway of plasma coagulation corroborated nM endogenous thrombin concentrations measured in porcine blood activated by endotoxin insult in vivo. The model fit to data suggests that only about 20% conversion of blood fibrinogen to fibrin is required to coagulate (gel) porcine plasma. This prediction is consistent with the common clinical laboratory observation of latent fibrin formation in "serum" separated from blood before fibrinogen is fully converted to fibrin. It was concluded that the thrombin-binding anticoagulation model was a reasonable simulation of the situation in which an initial bolus of either exogenous or endogenous thrombin is rapidly partitioned between fibrinogen-bound and ligand-bound forms with little or no additional free thrombin created over time.
Vogler
E A
EA
Becton Dickinson Research Center, Research Triangle Park, North Carolina 27709, USA. Vogler@bdrc.bd.com
Nadeau
J G
JG
Graper
J C
JC
D000818:Animals; D000925:Anticoagulants / Q000378:metabolism*; D001777:Blood Coagulation; D002417:Cattle; D005340:Fibrinogen / Q000378:metabolism; D008954:Models, Biological; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D013552:Swine; D013917:Thrombin / Q000378:metabolism*
D000925:Anticoagulants; D009838:Oligodeoxyribonucleotides; D005340:Fibrinogen; D013917:Thrombin