Thromboplastin immobilized on polystyrene surface exhibits kinetic characteristics close to those for the native protein and activates <Emphasis Type="Italic">in vitro</Emphasis> blood coagulation similarly to thromboplastin on fibroblasts |
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Authors: | O A Fadeeva M A Panteleev S S Karamzin A N Balandina I V Smirnov F I Ataullakhanov |
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Institution: | 1.National Research Center for Hematology,Russian Academy of Medical Sciences,Moscow,Russia;2.Center of Theoretical Problems of Physicochemical Pharmacology,Russian Academy of Sciences,Moscow,Russia;3.Physical Faculty,Lomonosov Moscow State University,Moscow,Russia |
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Abstract: | A method for transmembrane protein thromboplastin (tissue factor) immobilization on polystyrene surface is described. Tissue
factor is the main activating factor launching the blood coagulation process. It is a cofactor of factor VIIa, the first protease
in the cascade of coagulation reactions. The proposed method preserves kinetic characteristics specific for native tissue
factor on the fibroblast surface. The kinetics of binding to factor VIIa and enzymic activity of the formed complex follow
Michaelis-Menten kinetics, which is also characteristic of native complex. A small difference is that dissociation constant
for tissue factor immobilized on polystyrene surface exceeds 2.7-fold that for native factor. The proposed technique of immobilization
provides for protein density on the activating surface corresponding to the tissue factor density on the fibroblast surface.
The immobilized tissue factor can be used to activate blood coagulation in methods simulating spatial dynamics of in vitro clot growth. Investigation in this direction will make it possible to register both hypo- and hypercoagulation states of
the system. This approach is advantageous over traditional methods of estimation of the coagulation system conditions, which
mainly register only hypocoagulation. Investigation of the storage time has shown that activators containing immobilized tissue
factor can be stored and used during for at least 100 days in the method studying spatial dynamics of fibrin clot formation. |
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