Abstract
In search for the adequate assay method for the hypercoagulable state, the chromatographic method was introduced which revealed the development of macromolecular fibrinogen/fibrin complex in plasma of hypercoagulable state by the principle of gel exclusion chromatography. Plasma specimens of 16 normal subjects and 80 patients including thrombotic diseases were chromatographed on Biogel A-5m column (15×300mm) with 0.02M borate buffer containing 0.38% citrate (pH 7.5). The elution profiles of clottable fibrinogen, fibrinogen-related antigen (FR-ag) were obtained. The elution volumes of the beginning of FR-ag from the column were measured and these values were arbitrarily named as “initiation volumes (Vi's)”.
The elution profiles of FR-ag were classified into three: normal, thrombotic and DIC. Vi's of normal plasma and non-thrombotic diseases were the average of 20.5ml, but those of thrombotic diseases and DIC were 18.0ml with the significant difference from the former. These results suggested the development of macromolecular fibrinogen complex in the hypercoagulable states.
Reproduction of macromolecular complex was tried in vitro. The chromatography revealed the thrombotic profiles and the decrease of Vi's of plasma specimens such as normal plasma mixed with 0.04ml thrombin (f. c.), cryoprecipitate from the normal plasma mixed with 0.08u/ml thrombin (f. c.) and normal plasma mixed with soluble fibrin in urea. Only the FDP gave the DIC profile with the decreased Vi, which was prepared immediately after the complete dissolution of fibrin clot with plasmin. At the experiment in vivo, intravenous injection of 12μg/kg of Russel Viper venom into rabbits brought about the temporary shortening of PTT, but no significant change of PT, A-PTT, fibrinogen or platelet count. The elution volumes of fibrinogen of these plasmas remained unchanged, but Vi's after the injection were apparently decreased an the elution profiles of FR-ag formed DIC profiles. It was concluded that these experiments supported the results of the clinical specimens and the usefulness of this assay method for the hypercoagulable state including thrombosis and DIC.
