Abstract: Provided by the present invention are novel methods of factor IX protein recovery and purification.

Abstract: A thrombin of human or animal origin is free of infectious agents and is produced by activation of prothrombin subjected to a heat treatment for the inactivation of infectious agents.

Abstract: The present invention describes a method for controlled activation and degradation of FVII during purification whereby a solution of Factor VII is subjected to a number of chromatographic purification steps wherein Zn.sup.++ is present in at least one of the purification steps.The present invention also describes a method for controlled activation and degradation of FVII wherein a solution of FVII is applied to a number of anion exchange and immunoaffinity columns.

Abstract: The invention relates to a stable preparation which comprises a protein that is bound in and/or on lipid vesicles and that was treated for the inactivation of potentially present viruses. Further, the invention relates to methods for the production of a stable preparation for the treatment of blood coagulation disorders, wherein a protein is bound in and/or on lipid vesicles, and the method comprises a step in which the protein lipid complex is subjected to a treatment for the inactivation of potentially present viruses.

Abstract: A description is given of the possibility of using transglutaminases in a process for the preparation of an immunosuppressant. Additionally described is a pharmaceutical containing a transglutaminase and a plasminogen activator inhibitor.

Abstract: An improved assay for determining sensitivity to activated protein C in test samples has been developed to ensure rapid and accurate evaluations. This assay is based on measuring the conversion, by activated factor VIII within a test sample, of added factor X to an activated form. The activated protein C sensitivity of the test sample is determined by the relative inhibition of factor X conversion as compared to a control. A test sample that has decreased sensitivity to activated protein C will show relatively low inhibition, and vice versa.

Abstract: A method of recovering active, highly purified and concentrated vitamin K-dependent proteins from plasma, concentrate or mixtures of proteins produced by recombinant DNA technology using an immunoadsorbent comprising a monoclonal antibody, and the active, highly purified and concentrated vitamin K-dependent protein produced by the method.

Abstract: Analogs of Factor Xa (Factor Xai) which are inactive as proteases in the prothrombinase reaction are useful in treatment of diseases characterized by thrombosis. These antithrombotic agents can be conveniently prepared using recombinant techniques.

Abstract: A method for the separation of vitamin K-dependent proteins from non-vitamin K-dependent accompanying proteins is described wherein the method is characterized in that at least anion exchange chromatography and optionally affinity chromatography is carried out as well. The method is suitable especially for the purification of Factor II, VII, IX, X as well as Protein S, Protein C and Protein Z. With the aid of the method according to the invention a vitamin K-dependent protein is obtained which is present at a purity of 95%.

Abstract: A process for producing a highly purified preparation of an inhibited form of an activated blood factor entails providing a partially purified preparation containing the blood factor of interest, treating the partially purified preparation to convert the blood factor to an inhibited activated form in a single step, and then purifying the resulting inhibited activated blood factor.

Abstract: A method of making a platelet releasate product is disclosed involving performing an assay on a platelet releasate sample for a component of the releasate and forming platelet releasate product by comparing the assay results to a predetermined range of amounts of the component to be contained in the product. A method of treatment of tissue is disclosed involving the topical application of such product.

Abstract: Analogs of Factor Xa (Factor Xai) which are inactive as proteases in the prothrombinase reaction are useful in treatment of diseases characterized by thrombosis. These antithrombotic agents can be conveniently prepared using recombinant techniques.

Abstract: A process for producing a highly purified preparation of an inhibited form of an activated blood factor entails providing a partially purified preparation containing the blood factor of interest, treating the partially purified preparation to convert the blood factor to an inhibited activated form in a single step, and then purifying the resulting inhibited activated blood factor.

Abstract: A process for producing a highly purified preparation of an inhibited form of an activated blood factor entails providing a partially purified preparation containing the blood factor of interest, treating the partially purified preparation to convert the blood factor to an inhibited activated form in a single step, and then purifying the resulting inhibited activated blood factor.

Abstract: Analogs of blood factors which are transiently inactive are useful in treatment of diseases characterized by thrombosis. In addition, modified forms of activated blood factors that generate the active blood factor in serum but have extended half-lives are useful in treating hemophilic conditions. These modified forms of the blood factor may be acylated forms which are slowly deacylated in vivo.

Abstract: This invention relates to methods of removing undesired antibodies from blood-derived compositions containing both the antibodies and coagulation factors, such that the coagulation factors are substantially retained in the composition. The undesired antibodies may be blood group antibodies. This invention also relates to compositions in which undesired antibodies have been removed and desired coagulation factors are retained. This invention further relates to methods of inactivating virus and removing undesired antibodies from blood-derived compositions containing virus, antibodies and coagulation factors without removing coagulation factors therefrom, and to the resulting compositions.

Abstract: There is disclosed the use of prothrombin fragments, preferably of human prothrombin fragments, having a thrombin-like activity, in particular of meizothrombin, meizo thrombin (desF1), or mixtures thereof, for diagnostic purposes for assaying thrombin substrates as well as a reagent containing these prothrombin fragments.

Abstract: Assay methods for diagnosing blood clotting disorders are described. The assays use data bases for pooled normal plasma (PNP) and plasma from healthy volunteers, males and females ages 18 to 64 years. Charting on a comparative basis of patient plasma and PNP allows the results to be interpreted by reference to the data base. Simple, rapid, inexpensive and highly sensitive and specific assays devised for diagnosing blood clotting disorders are described.

Abstract: To determine the factor VIII activity of a sample, the latter is reacted with a reagent containing thrombin, factor IXa.beta., phospholipids and calcium ions so as to form a factor VIIIa-factor IXa-phospholipid-calicum ion-containing complex. This complex is reacted with factor X so as to obtain activated factor X. The obtained factor Xa is reacted with chromogenic substrate so as to form a substance to be determined spectrophotometrically. Complex formation and activation of factor X are effected in one stage: The sample is mixed with a mixture of human thrombin, phospholipids, calcium ions, human factor IXa.beta., human factor X and, if desired, human factor XIa and human factor XIIa, as a complex-forming reagent.

Abstract: It has been discovered that it is possible to administer truncated tissue factor, not having the transmembrane region (tTF) in combination with factor VIIa (FVIIa) to treat bleeding disorders such as those resulting from hemophilia or cirrhosis of the liver. Preferably, the tTF is administered to produce up to 10 .mu.g tTF/ml of plasma. The FVIIa is administered to produce levels of between 40 ng FVIIa/ml and 4 .mu.g FVIIa/ml of plasma. The effective dosages of both tTF and FVIIa are significantly and surprisingly less than the administration of either alone to stop bleeding. Examples demonstrate safety and efficacy in normal and hemophilic dogs.

Abstract: It has been discovered that it is possible to administer truncated tissue factor (not having the transmembrane region) (tTF) in combination with factor VIIa (FVIIa) or an activator of endogenous factor VII to treat bleeding disorders such as those resulting from hemophilia or cirrhosis of the liver. The tTF is administered to produce up to 10 .mu.g tTF/ml of plasma. The FVIIa or FVII activator is administered to produce levels of between 40 ng FVIIa/ml and 700 ng FVIIa/ml of plasma. The effective dosages of both tTF and FVIIa/factor VII activator are significantly and surprisingly less than the administration of either alone to stop bleeding. Examples demonstrate safety and efficacy in normal and hemophilic dogs.

Abstract: An assay for activated factor VII (factor VIIa) has been developed using truncated tissue factor (tTF), a soluble mutant form of (tTF) that retains the cofactor function of TF toward factor VIIa. Unlike full-length TF, however, tTF appears not to support the conversion of factor VII to VIIa. As a result, the tTF assay for factor VIIa is free from interference from factor VII in the plasma and is therefore specific for factor VIIa. The assay is much simpler than existing assays, because it is a single-stage clotting assay performed almost identically to a prothrombin time assay. It is also considerably more sensitive than current assays for factor VIIa in plasma. Since the tTF assay is calibrated against a factor VIIa standard, it yields an absolute concentration of factor VIIa in ng/ml.

Abstract: A description is given of the possibility of using transglutaminases in a process for the preparation of an immunosuppressant.Additionally described is a pharmaceutical containing a transglutaminase and a plasminogen activator inhibitor.

Abstract: The present invention relates to a method for preparing a high purity Factor IX concentrate. The starting material for the process is the supernatant fraction of a cryoprecipitated human plasma. A pre-purification step is performed by DEAE-Sephadex chromatography. The resulting Factor IX fraction has a specific activity of at least 0.5 IU/mg protein. The purification method of the invention comprises two successive chromatography separations. First, ion-exchange chromatography on DEAE-sepharose is conducted so that the Factor IX is eluted when the ionic force of the buffer is increased to 0.34-0.38M sodium chloride. Then, affinity chromatography is conducted on heparin-sepharose. The elution buffer is a citrate buffer at a pH of 7.4 adjusted with 0.45M sodium chloride and supplemented with arginine as a stabilizer for Factor IX activity. Lysine is added as a stabilizer before freeze-drying of the purified Factor IX.

Abstract: The present invention provides isolated DNA molecules comprising a DNA segment encoding novel human Kunitz-type inhibitors. Also provided are DNA constructs comprising a first DNA segment encoding a novel human Kunitz-type inhibitor wherein said first DNA segment is operably linked to additional DNA segments required for the expression for the first DNA segment, as well as host cells containing such DNA constructs and methods for producing proteins from the host cells.

Abstract: A method for at least partially separating vitamin K-dependent blood clotting factors from a mixture containing at least one such factor, e.g. a prothrombin complex concentrate, which comprises adsorption of said mixture on to a chelate of a polyvalent metal immobilized on an inert support, e.g. Cu.sup.2 + -primed Chelating Sepharose, followed by elution to yield one or more fractions enriched in respect of one of said factors.

Abstract: A dry reagent consisting essentially of (a) a protein having thrombin activity, (b) at least one additive selected from the group consisting of amino acid, a salt thereof and saccharide and optionally (c) magnetic particles, and a method of assaying fibrinogen in an assay sample using the above dry reagent. The dry reagent obviates the time required for preparing a reagent and warming an assay sample, and permits facile fibrinogen assay by only diluting an assay sample. The fibrinogen assay range being broad, the dry reagent substantially obviates the procedure of remeasuring plasma having a fibrinogen concentration outside the assay range of a liquid reagent. The assay result by the dry reagent and that by a liquid reagent well correlate with each other as compared with the result by any known thrombin-containing dry reagent, and the assay by the dry reagent can be performed with good reproducibility in achievement and reliability in measurement.

Abstract: The subject invention provides a pharmaceutical composition, which comprises an amount of calreticulin effective for blocking or preventing thrombosis in a subject, causing substantially no defect or no defect in normal hemostasis, and a pharmaceutically effective carrier. The subject invention provides a method for blocking or preventing thrombosis in a subject, causing substantially no defect or no defect in normal hemostasis, which method comprises administering calreticulin to the subject in an amount effective for blocking or preventing thrombosis. The subject invention provides a pharmaceutical composition, which comprises calreticulin in combination with an other antithrombotic agent, in an amount and proportion effective for enhancing the action of the other antithrombotic agent, to prevent clotting or dissolve clots which have already formed.

Abstract: The Prothrombin Time (PT) is used as a screening test for blood coagulation factor deficiencies and for monitoring oral anti-coagulant therapy using coumadin. Thromboplastin reagents activate the extrinsic pathway of coagulation and are the basis for the Prothrombin Time (PT) test. This invention describes the use of barium sulfate and chaotropic agents, and nonionic detergents, for the extraction of sensitive thromboplastin reagents from tissue. Extraction with sodium thiocyanate alone also greatly enhances thromboplastin sensitivity. This invention should be useful for all thromboplastins and will improve their sensitivity for all PT-based tests and specific assays.

Abstract: Test articles comprise a solid phase membrane having dry thromboplastin immobilized thereon or within. The thromboplastin is substantially free from substances which might cause aberrant functioning intermediate transition states as the thromboplastin is rehydrated with liquid sample. Coagulation neutral agents which facilitate rehydration of the dry thromboplastin are also provided on the solid phase membrane.

Abstract: A reagent blood coagulation that causes an increase in turbidity changes when added to a plasma sample containing a substance activating coagulation factor activating such as tissue thromboplastin, phospholipid and thrombin, calcium ion, and molecular substance such as high molecular vinyl and a high molecular polysaccharide. By adding a molecular substance to the reagent, the turbidity change due to blood coagulation is increased, and hence the changing quantity of transmitted light or scattered light increases, thereby making it possible to achieve a more accurate detection. Besides, by adjusting the electric conductivity, pH and osmotic pressure to proper values, the turbidity change due to blood coagulation may be further amplified.

Abstract: An ultra-pure, clear thrombin solution having a high specific activity is described as well as a method of manufacture.

Abstract: The present invention relates to a process of making a concentrate of coagulation proteins starting with whole human or animal plasma. This concentrate is used as a biological adhesive when extemporaneously mixed to thrombin. The concentrated proteins include mostly fibrinogen, fibrin stabilizing factor (factor XIII) and fibronectin. The claimed process has the advantage of being short of execution while providing an excellent yield of coagulable proteins. No protease inhibitor has to be added during the process. The process involves steps of separation by "salting-out" in presence of amino-6 hexanoic acid which prevents co-precipitation of plasminogen with the desired coagulable proteins. The proteins so obtained are very stable after reconstitution in water for at least 24 hours at room or body temperature. After mixing with thrombin and calcium, the adhesive shows excellent strength and biocompatibility.

Abstract: A description is given of a process for the purification of factor IX, factor X, and factor II from human plasma or fractions thereof, wherein a solution containing prothrombin complex factors is purified by repeated ion exchange chromatographic separations followed by adsorption chromatography on metal ions.

Abstract: A pure Factor I protein essentially free of infectious virus. Factor B and C3. The protein is derived from plasma and pasteurized by heating to a temperature of 50.degree. to 65.degree. C. for 0.5 to 100 hours in the presence of one or more stabilizers for Factor 1. Preparations containing the protein are useful in the treatment of Factor I deficiency and autoimmune diseases.

Abstract: Use of human blood coagulation factor XIII for the treatment of ulceration colitisThe use of human blood coagulation factor XIII for the treatment of ulcerative colitis is demonstrated in 4 representative cases. Treatment with factor XIII leads to rapid and total disappearence of the chief symptoms realizing transition to the remission stage.

Abstract: It has been discovered that it is possible to administer truncated tissue factor (not having the transmembrane region) (tTF) in combination with factor VIIa (F VIIa) to treat bleeding disorders such as those resulting from hemophilia or cirrhosis of the liver. The tTF is administered to produce up to 10 .mu.g tTF/ml of plasma. The F VIIa is administered to produce levels of between 40 ng VIIa/ml and 4 .mu.g F VIIa/ml of plasma. The effective dosages of both tTF and VIIa are significantly and surprisingly less than the administration of either alone to stop bleeding. Examples demonstrate safety and efficacy in normal and hemophilic dogs.

Abstract: In the method for purifying factor VIII from cryoprecipitate, which is dissolved and then treated with alumina gel, the extract is diluted to a protein concentration not exceeding approximately 5 g/l and subjected to viral inactivation with solvent/detergent, the inactivated extract containing the solvent/detergent is then subjected to chromatography on a weak anion exchange column which is hydrophilic in nature and factor VIII is then eluted with a dissociating buffer.

Abstract: A purified therapeutic grade thrombin is described which is essentially free of lipid envelope viruses, has a specific activity of about 2200 NIH units per milligram of protein to about 3200 NIH units per milligram of protein, is essentially homogeneous and may be produced on a commercial-scale. The thrombin is acceptable from human administration.

Abstract: The invention relates to a process for the manufacture of a high-purity activated factor VII concentrate.This process comprises the use of a plasma free of cryoprecipitate, preferably of human origin, as well as at least one purification step involving chromatography at least once on an ion exchange resin, and a factor VII activation step, wherein the first stage is direct activation of the factor VII in the crude supernatant of plasma free of cryoprecipitate, without the addition of exogenous proteins.By virtue of the invention, the high-purity activated factor VII is essentially free of vitamin K-dependent factors and factors VIIIC and VIIICAg and has a factor VIIa/factor VII ratio greater than 5 with a specific activity of the activated factor VII greater than 200 IU/mg of proteins.

Abstract: An apparatus for resisting coagulation of blood, includes a blood container having disposed therein antibody to blood coagulation factors for resisting coagulation of blood. The apparatus further includes a needle and a holder for the needle which are both operatively associated with the blood container. The blood coagulation factor antibody inhibits coagulation of the blood drawn by the needle into the blood container so that an uncoagulated blood sample is obtained.

Abstract: The present invention provides a fail-safe combination of chemical and physical means for rendering a blood product which comprises a labile blood protein free of viruses without incurring protein denaturation. The blood product is contacted with an effective amount of a selected chemical disinfectant, preferably, sodium thiocyanate in combination with a physical process, preferably, ultrafiltration. The blood product may be plasma, serum, plasma concentrate, cryoprecipitate, cryosupernatant, plasma fractionation precipitate or plasma fractionation supernatant containing viruses such as hepatitis or human immunodeficiency virus.

Abstract: The present invention relates to a process for purifying Factor IX from an impure protein fraction containing Factor IX. The purification process comprises the steps of adding a solvent and a detergent to an impure protein fraction and incubating the solvent/detergent protein solution to inactivate any viral contaminants. Factor IX is purified from the solvent/detergent protein solution by chromatography on a sulfated polysaccharide resin without first removing the solvent and detergent prior to the purification on the sulfated polysaccharide resin. The Factor IX, purified by the process has a specific activity of at least 85 units/mg.

Abstract: The present invention describes APC polypeptides and anti-peptide antibodies capable of inhibiting activated Protein C anticoagulant activity. The polypeptide and antibody are useful in diagnostic methods and systems for measuring APC in vascular fluid samples. In addition, the polypeptide and anti-peptide antibody are useful in therapeutic methods for inhibiting APC in a human patient.

Abstract: Analogs of Factor Xa (Factor Xai) which are inactive as proteases in the prothrombinase reaction are useful in treatment of diseases characterized by thrombosis. These antithrombotic agents can be conveniently prepared using recombinant techniques.

Abstract: The Prothrombin Time (PT) is used as a screening test for blood coagulation factor deficiencies and for monitoring oral anti-coagulant therapy using coumadin. Thromboplastin reagents activate the extrinsic pathway of coagulation and are the basis for the Prothrombin Time (PT) test. This invention describes the use of barium sulfate and chaotropic agents, and nonionic detergents, for the extraction of sensitive thromboplastin reagents from tissue. Extraction with sodium thiocyanate alone also greatly enhances thromboplastin sensitivity. This invention should be useful for all thromboplastins and will improve their sensitivity for all PT-based tests and specific assays.

Abstract: A method for the recombinant production of zymogen forms of human protein C is described. These zymogen forms differ from native zymogen protein C in their increased sensitivity to activation by thrombin and thrombin/thrombomodulin. DNA compounds, vectors, and transformants useful in the method are also disclosed.

Abstract: Proteins which specifically inhibit coagulation Factor Xa. The inhibitors, which do not inhibit Factor VIIa, kallikrein, trypsin, chymotrypsin, thrombin, urokinase, tissue plasminogen activator, plasmin, elastase, Factor XIa or S. aureus V8 protease, are polypeptides of 60 amino acid residues. The inhibitors may be purified from Ornithodoros moubata extract, synthesized, or produced using a recombinant DNA yeast expression system.

Abstract: Methods are disclosed for producing hybrid phospholipid-binding proteins from eukaryotic cells. DNA constructs comprising a transcriptional promoter, at least one signal sequence and a hybrid phospholipid-binding protein coding sequence comprising at least one lipocortin lipid-binding domain joined to a gla-domainless, vitamin K-dependent protein and a transcriptional terminator are also disclosed.

Abstract: The present invention describes a process for activating Factor II to Factor II.sub.a by incubating Factor II in the presence of Factor V, Factor X.sub.a, phospholipids, and calcium ions. Each of the factors is prepared from a single impure protein fraction which includes Factors II, V and X. The Factor II, V and X purification procedure comprises the steps of DEAE ligand chromatography and precipitation by the addition of barium chloride. Factor V is recovered from the barium chloride supernatant, and Factors II and X are contained in the barium chloride precipitate. The barium chloride precipitate is dissolved in an aqueous solution and is applied to a chromatographic resin coupled with a ligand which binds Factor X and Factor II weakly or not at all. Factor II is recovered from the fraction, which remains unbound or weakly bound to the Factor X binding ligand. Factor X.sub.