Concentrates of coagulation factors II, VII, IX and X, method of their preparation and use
AssigneeTessek sdruzeni Praha
InventorsMiroslav Rybak, Evzen Kasafirek, Jitka Houskova, Cyril Losticky, Stanislav Ulrych, Oldrich Sedlmaier, Alena Roubalova
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Abstract
The claimed invention concerns concentrates of coagulation factors II, VII, IX and X and the method of their preparation by selective adsorption on an equilibrated carrier of hydroxyethylmethacrylate or hydroxyethylacrylate with bound diethylaminoethyl groups, followed by selective elution. The concentrates of coagulation factors II and X can be used to detect coagulation factor VII and in a laboratory method for the detection or determination of the extent of mammalian inflammatory diseases.
Claims14
A concentrate of a coagulation factor selected from the group consisting of coagulation factors II, VII, IX and X, wherein the coagulation factor is prepared by contacting a biological material selected from the group consisting of human blood plasma, animal blood plasma and fractions thereof, containing one or more of the above mentioned factors, with a sorbent based upon a synthetic matrix selected from the group consisting of polymers of diethylaminoethyl-hydroxyethylacrylate, polymers of diethylaminoethyl-hydroxyethylmethacrylate, and mixtures thereof, and desorbing the desired coagulation factor by eluting the sorbent with a buffered solvent having a pH from 7.2-7.6 and a saline content of 0.3-2.0 mol/l.
A method for detecting the presence of factor VII in a fluid comprising contacting said fluid with concentrates of coagulation factors II and X in the presence of thromboplastin, phospholipids, Ca.sup.2+ -ions and a chromogenic substrate, and observing whether a color change occurs, indicating the presence of factor VII.
A method of preparation of coagulation factor concentrates selected from the group consisting of coagulation factors II, VII, IX and X comprising contacting a human or animal blood plasma or a fraction thereof, wherein said fluid is contacted with a sorbent based upon a synthetic matrix selected from the group consisting of polymers of diethylaminoethyl-hydroxyethylacrylate, polymers of diethylaminoethyl-hydroxyethylmethacrylate, and mixtures thereof, equilibrated with a buffer with a pH of 7.2-7.6 and a saline concentration of 0.1 mol/l, removing the unbound proteins by washing with the same buffer containing 0.2 mol/l saline, and desorbing the desired coagulation factors with a buffer containing saline at a concentration from 0.3 to 2.0 mol/l.
A method for determination of the presence of coagulation factor VII in a biological fluid for the determination of the extent of mammalian inflammatory disease by analysis of the body fluids wherein samples of said biological fluid are collected and then are simultaneously or in a stepwise manner brought into contact with thromboplastin, phospholipids, concentrate of coagulation factors II and X, Ca.sup.2+ -ions and a chromogenic substrate and evaluating the resulting color.
Dependencies are inferred from the claim text.