The use of fluorogenic substrates to monitor thrombin generation for the analysis of plasma and whole blood coagulation.
Thrombin is central to the process of coagulation and monitoring its activity is a reliable indicator of the rate and extent of coagulation. I have employed a range of fluorogenic peptide substrates as indicators of coagulation via the formation of active thrombin. This system enabled coagulation to be monitored in a kinetic fashion, and the use of fluorescence enabled a wide range of samples to be analyzed including lyophilized plasma containing fibrin, fresh platelet-poor plasma, platelet-rich plasma, and even whole blood. Coagulation could be monitored following triggering by tissue factor, ellagic acid, or each of the proteases preceding thrombin in the coagulation network. Using this assay procedure I have investigated the anticoagulant activities of a number of compounds and the results indicate that this assay would be useful for the kinetic analysis of coagulation in various plasma preparations, or even whole blood.
Ramjee
M K
MK
Peptide Therapeutics Group plc., Peterhouse Technology Park, Cambridge, CB1 9PT, United Kingdom.
D001777:Blood Coagulation / Q000187:drug effects; D001780:Blood Coagulation Tests; D004610:Ellagic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D005456:Fluorescent Dyes; D006801:Humans; D007202:Indicators and Reagents; D007700:Kinetics; D008991:Monitoring, Physiologic / Q000379:methods; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D015203:Reproducibility of Results; D013917:Thrombin / Q000032:analysis* / Q000378:metabolism; D013925:Thromboplastin / Q000378:metabolism
D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D011994:Recombinant Proteins; D004610:Ellagic Acid; D013925:Thromboplastin; D010450:Endopeptidases; D013917:Thrombin