Abstract: Methods of treating neuroinflammation by administration of a selective protein C activator, such as recombinant human WE thrombin and optionally one or more of its cofactors are disclosed. Also disclosed are pharmaceutical compositions for use in the treatment of mammals that exhibit symptoms of neurological inflammation. The pharmaceutical compositions and pharmacological dose comprise a safe and effective amount of WE thrombin.

Abstract: The present invention provides methods for determining whether activated carbon can be used for removing viruses or a certain virus from a sample containing a target protein.

Abstract: Compositions comprising extracellular matrix (ECM) materials and methods of use thereof are disclosed. The compositions may comprise two or more ECM materials derived from different types of tissues, such as, e.g., lung tissue and spleen tissue, formulated for administration to a patient or configured as a medical device for implantation in or application to the patient. The compositions may combine complementary properties of different types of ECM materials for customized patient-specific and/or site-specific tissue repair and/or regeneration.

Abstract: Provided are methods for lyophilization of an aqueous thrombin solution, thrombin solutions for use in such lyophilization methods, and solid thrombin compositions produced by such methods.

Abstract: A process for preparing a bioabsorbable sheet preparation holding thrombin is provided. A process for preparing a bioabsorbable sheet preparation holding thrombin which comprises immersing a bioabsorbable sheet consisting of polyglycolic acid in a thrombin solution containing thrombin as an active ingredient, glycerol as a softening agent, Tween 80 as a permeating agent, and optionally histidine and trehalose as a stabilizing agent followed by drying to hold thrombin on said bioabsorbable sheet, and a bioabsorbable sheet preparation holding thrombin prepared by said process.

Abstract: Methods, apparatus, and compositions related to generating and using thrombin. Methods include preparing a solution comprising thrombin by precipitating fibrinogen from a liquid comprising whole blood or a blood fraction. Precipitated fibrinogen is removed from the liquid to form a post-precipitation liquid that is incubated with calcium and a plurality of beads to form a clot. A solution comprising thrombin is separated from the clot. Thrombin prepared thereby can be used as a tissue sealant and in methods of applying a tissue sealant to subject, including application of an autologous tissue sealant.

Abstract: One aspect of the invention contemplates a mutant E-WE thrombin precursor that contains the SEQ ID NO:1 amino acid residue sequence. Another aspect contemplates a thrombin precursor that contains the amino acid residue sequence Asp/Glu-Gly-Arg at positions 325, 326 and 327 based on the preprothrombin sequence. A third aspect contemplates a thrombin precursor that contains the SEQ ID NO:1 amino acid residue sequence as well as the amino acid residue sequence Asp/Glu Gly Arg at positions 325, 326 and 327 based on the preprothrombin sequence. Also contemplated is a composition that contains an effective amount of mutant thrombin dissolved or dispersed in a pharmaceutically acceptable carrier. A method is also disclosed for enhancing treating and preventing thrombosis in a mammal in need using that composition.

Abstract: The present invention relates to a chromogenic method for simultaneously determining the activity of multiple coagulation proteases or for simultaneously determining the inhibition of multiple coagulation proteases in a single test reaction. For this purpose, use is made of two chromogenic substrates which have different absorption maxima and whose color signals can be separated spectrally.

Abstract: This document relates to compositions containing cardiogenic factors, to methods to obtain cells by culturing initial cells in the presence of such factors; and methods of administering the obtained cells to heart tissue.

Abstract: The present invention disclosed the cloning and the purification of a mutant of prethrombin-2, which contains a thrombin cleavage site instead of a factor Xa cleavage site. The stable mutant prethrombin-2 is able to convert itself autocatalytically into active ?-thrombin in the absence of ecarin or factor Xa. The new concept of signal amplification using self-replicating enzymes can be applied to improve sensitivity of ?-thrombin assays and also for the preparation of different enzymes.

Abstract: The present invention provides a recombinant baculovirus. The baculovirus has a genome into which a gene encoding ?-glutamyl carboxylase (GGCX) and a gene encoding DT-diaphorase (NQO1) are incorporated. The present invention further provides a method for producing a recombinant vitamin K-dependent protein by using the recombinant baculovirus.

Abstract: The present invention provides a method for producing recombinant prothrombin. The method comprises: providing a vector DNA into which a gene encoding a tag and a gene encoding prothrombin are incorporated, wherein the tag is selected from the group consisting of MBP, SUMO, and NusA; and expressing a tag fusion type prothrombin in a lepidopteran insect or cultured cells of the lepidopteran insect.

Abstract: The invention relates to a thrombolytic enzyme referred to as Thrombinase having a molecular weight of 31,000 to 32,000. Such a thrombolytic enzyme can be used for dissolving blood clots. The process comprises culturing a filtrate of Bacillus sphaericus sero type H5a 5b, removing the cell, subjecting the cell supernatant to filtration, salting out the retentate, subjecting the precipitate to dialysis, reprecipitating the precipitate and then reconstituting in buffer and finally decolorizing, purifying and dialyzing.

Abstract: Provided is a hemostasis assay comprising a reaction mixture comprising a blood product to be tested, a trigger molecule for inducing thrombin generation, a thrombin-specific substrate which, upon cleavage by thrombin, produces a measurable thrombin-specific signal, a trigger molecule for inducing plasmin generation, a plasmin-specific substrate which, upon cleavage by plasmin, produces a measurable plasmin-specific signal, a phospholipid-containing surface, and calcium ions. The assay allows determination of the amount of thrombin and the amount of plasmin generated in the reaction mixture in time, starting at t=0, by measuring the thrombin-specific and plasmin-specific signals.

Abstract: A method is provided for determining in real time the course of thrombin activity in a sample of blood or plasma as it appears in and disappears from the simple which comprises adding a thrombin substrate to the sample that, per unit time, produces a detectable signal in a quantity that bears relation to the amount of thrombin present. Simultaneously, in a control sample of the same blood or plasma in which thrombin generation is not triggered, the activity of a standard preparation with invariable thrombin activity is measured. The exact molar amount of thrombin present at any moment is obtained by comparison of the activity measured in clotting blood and the simultaneously measured calibrator. The method is useful inter alia for diagnosing hyper- and hypo-coaguable states, either congenital, acquired or drug-induced in humans and animals. Also provided is a kit for use in this method.

Abstract: A method of alleviating pain associated with tissue damage includes applying salmon thrombin at a tissue damage site, as a single substance in liquid form, or as a powder, a foam, and/or a gel that includes salmon thrombin. A pain relief substance includes a salmon thrombin preparation.

Abstract: Methods of generating thrombin and methods of applying a clotting tissue sealant to a site on a subject are provided. A blood component comprising platelets can be obtained from the subject. A hypotonic composition is contacted with a solid matrix to form a thrombin-containing liquid, where the hypotonic composition includes water, calcium, a blood component comprising platelets, and optionally a chelator. Calcium is present in the hypotonic composition in an amount greater than the amount of calcium that can be complexed by the chelator. Thrombin-containing liquid is then separated from the hypotonic composition and can be applied to the site on the subject to form a clot, for example, by combination with fibrinogen.

Abstract: To provide a method for stabilizing unstable ?-thrombin in a thrombin-containing solution, a solution containing stabilized ?-thrombin, and a liquid fibrinogen assay reagent containing the solution. The method for stabilizing ?-thrombin in a thrombin-containing solution, which includes adjusting the percentage of ?-thrombin to 70% or more with respect to the amount of total thrombin in the thrombin-containing solution.

Abstract: A composition is provided, which comprises a serine protease; a reversible inhibitor of said serine protease; and a stabilizing agent M having the formula I: Also provided are uses of the composition as a medicament, and other uses and methods employing its various properties.

Abstract: A method of detecting an abnormal amount of a biomarker associated with biliary tract cancer in a subject can comprise: a) quantitating an amount of a fragment of prothrombin having an m/z value of about 4204 m/z in a biological sample from the subject; and b) comparing the quantitated value obtained in (a) with a threshold value.

Abstract: Novel thrombin/prothrombin protease/zymogen variants which have anticoagulation activity and methods of use thereof are disclosed.

Abstract: The invention features methods and compositions for assessing risk, particularly immediate risk, of thrombotic events in patients with suspected or known vascular disease, and more particularly to assessing risk of thrombotic events in patients with coronary artery disease, particularly acute myocardial infarction, stroke, unstable angina, stable angina, or restenosis. Risk of thrombosis can be assessed by analysis of platelet reactivity and/or velocity of thrombin or fibrin formation, and determining whether the patient has a score associated above a risk threshold value. In other embodiments, risk of thrombosis in a patient is evaluated in the context of a profile generated from values obtained from one or more assays that evaluate various factors associated with thrombosis and/or atherosclerosis.

Abstract: The present invention relates to processes for the preparation of virus inactivated thrombin, and to the products prepared by these processes.

Abstract: A composition is provided, which comprises a serine protease; a reversible inhibitor of said serine protease; and a stabilizing agent M having the formula I: Also provided are uses of the composition as a medicament, and other uses and methods employing its various properties.

Abstract: The invention relates to a method for producing folded prethrombin, wherein inclusion bodies, which contain unfolded prethrombin or a derivative thereof, are solubilized in a solubilization buffer containing at least one chaotropic compound and at least one organic disulfide compound. The invention further relates to methods for producing thrombin and a-thrombin and derivatives thereof. The invention also relates to solutions that contain folded proteins, which can be produced by the methods according to the invention.

Abstract: The present invention provides polypeptides comprising or consisting of an amino acid sequence from thrombin, or a fragment, variant, fusion or derivative thereof, or a fusion of said fragment, variant or derivative thereof, for use in the treatment or prevention of inflammation and/or excessive coagulation of the blood. Related aspects of the invention provide isolated polypeptides comprising or consisting of an amino acid sequence of any one of SEQ ID NOs: 1 to 7, or a fragment, variant, fusion or derivative thereof, or a fusion of said fragment, variant or derivative thereof, which exhibit an anti-inflammatory activity, together with isolated nucleic acid molecules, vectors and host cells for making the same. Additionally provided are pharmaceutical compositions comprising a polypeptide of the invention, as well as methods of use of the same in the treatment and/or prevention of inflammation and/or excessive coagulation.

Abstract: A clotting serum production device including a main chamber, an inlet housing, and an outlet port. The main chamber is defined by a first end wall, a second end wall opposite to the first end wall, and a side wall extending between the first end wall and the second end wall. The inlet housing extends from the first end wall. The inlet housing includes a first inlet port, a second inlet port, and a first conduit extending between the first inlet port and a second conduit. The second conduit extends between the second inlet port and the main chamber. The inlet housing is configured to provide one-way fluid flow from the first inlet port to the second inlet port and to provide one-way fluid flow from the second inlet port to the main chamber. An outlet port is in fluid communication with the main chamber and provides fluid communication with an exterior of the clotting serum production device.

Abstract: The present invention relates to a batroxobin-encoding nucleotide sequence and/or a mutated ?-factor secretion signal sequence, and a vector and a transformant using the same. The batroxobin-encoding nucleotide sequence of this invention exhibits an excellent expression efficiency in yeast, particular Pichia pastoris and the recombinant batroxobin is obtained at 4-13 fold higher yield than natural-occurring batroxobin-encoding sequences. The protein expression system which uses the batroxobin-encoding nucleotide sequence as well as mutated ?-factor secretion signal peptide sequence of this invention obtains the recombinant batroxobin at about 20-fold higher yield than natural-occurring batroxobin-encoding sequences. In addition, the recombinant batroxobin prepared using the sequence of this invention has a significantly plausible activity and stability compared with natural-occurring batroxobin.

Abstract: A chimeric therapeutic polypeptide of a pre-existing therapeutic polypeptide is disclosed, as are a method of enhancing folded stabilization and a pharmaceutical composition of the glycosylated chimer. The pre-existing and chimeric polypeptides have substantially the same length, substantially the same amino acid residue sequence, and exhibit at least one tight turn containing a sequence of four to about seven amino acid residues in which at least two amino acid side chains extend on the same side of the tight turn and are within less than about 7 ? of each other. The chimeric therapeutic polypeptide has the sequon Aro-(Xxx)n-(Zzz)p-Asn-Yyy-Thr/Ser (SEQ ID NO:001) within that tight turn sequence such that the side chains of the Aro, Asn and Thr/Ser amino acid residues project on the same side of the turn and are within less than about 7 ? of each other. That sequon is absent from the pre-existing therapeutic polypeptide.

Abstract: The present invention discloses methods for the preparation of recombinant human proteins expressed in human cells. The present invention relates to methods for the preparation of human recombinant thrombin and human recombinant fibrinogen. The method employs serum-free culturing conditions and provides recombinant human proteins expressed in human cells of increased safety to the patient when used in human medical treatments. The immunogenic response to the recombinant human proteins expressed in human cells may be lower. Human recombinant thrombin is expressed in the human embryonic kidney 293 cell line and the protein can be prepared using two different routes, one starting from a point mutated prothrombin with gla and Kringle 1 and 2 domains, and maintaining these domains during the process; the other one starting with prothrombin (non-mutated), via a prethrombin with a HPC4-Kringle 2 domain and subjecting this prethrombin to a point mutation.

Abstract: An object of the present invention is to detect a small biomolecule in a sample using simple and inexpensive equipment. To achieve this, the small biomolecule in a sample S is detected optically. Specifically, the sample S containing an aptamer capable of interacting with the small biomolecule is irradiated with an excitation light Le and irradiated with a measurement light L2 for measuring the photothermal effect produced in the sample S by the irradiation with the excitation light Le. The photothermal effect induced in the sample S by the excitation light Le is measured from the phase change in the measurement light L2, and the presence or absence of the interaction between the biomolecule and the aptamer is assessed based on the temporal variation in the measurement signal.

Abstract: Methods, apparatus, and compositions related to generating and using thrombin. Methods include preparing a solution comprising thrombin by precipitating fibrinogen from a liquid comprising whole blood or a blood fraction. Precipitated fibrinogen is removed from the liquid to form a post-precipitation liquid that is incubated with calcium and a plurality of beads to form a clot. A solution comprising thrombin is separated from the clot. Thrombin prepared thereby can be used as a tissue sealant and in methods of applying a tissue sealant to subject, including application of an autologous tissue sealant.

Abstract: The invention relates to methods for determining the activity of a proteolytic coagulation factor of the blood coagulation cascade in a body fluid such as whole blood or plasma. A combination is provided in a reaction mixture. The combination comprises the sample and an activation agent for activating a proteolytic coagulation factor of the blood coagulation cascade or for activating the blood coagulation cascade. The effect of the activating on a reagent system comprising a cleavable moiety is evaluated. The cleavable moiety is or becomes bound to a chemiluminescent agent or a sensitizer agent or both. The chemiluminescent agent and the sensitizer agent are related in that, when in close proximity, energization of the sensitizer agent results in energization of the chemiluminescent agent. The effect of the activating is related to the activity of a proteolytic coagulation factor of the blood coagulation cascade wherein the effect is the extent of cleavage of the cleavable moiety.

Abstract: Methods for producing cell lines with high levels of biologically active recombinant vitamin K dependent proteins are described. The transfected cell lines do not include heterologous genes for processing enzymes and are not subject to selection pressure such as methotrexate resistance. Cell lines producing Factor VII/VIIa and Factor IX are described. These cell lines can be used for isolation of Factor VII/VIIa and/or Factor IX for treatment of Hemophilia.

Abstract: Methods of harvesting proteins directly from bioreactors to avoid at several steps in the purification of recombinant drugs are disclosed.

Abstract: The present invention relates to a method is provided for producing recombinant human thrombin from recombinant prothrombin using recombinant ecarin having the sequence SEQ ID NO 2 or a homologue thereof.

Abstract: A septum is positioned within a disposable vessel and defines a lower chamber and an upper chamber. The septum includes a plurality of apertures that provide fluid communication between the upper chamber and lower chamber. Compressed gas is introduced in the lower chamber to produce fine bubbles rising up throughout the vessel to produce a mixing and gasification needed for the growth of a biological culture and manufacture of a biological product in a nutrient medium. Adding a binding resin to the upper chamber allows harvesting, separation and purification of biological products in the reactor as a single unit operation.

Abstract: The present disclosure relates to a method for producing heterologous protein including: (c) providing a host cell comprising a 2 ?m-family plasmid, the plasmid comprising a gene encoding a protein comprising the sequence of a chaperone protein and a gene encoding a heterologous protein; (d) culturing the host cell in a culture medium under conditions that allow the expression of the gene encoding the chaperone protein and the gene encoding a heterologous protein; (e) purifying the thus expressed heterologous protein from the cultured host cell or the culture medium.

Abstract: Methods for producing proteins and glycoproteins in Pichia pastoris that lack detectable cross binding activity to antibodies made against host cell antigens are described. In particular, methods are described wherein recombinant Pichia pastoris strains that do not display a ?-mannosyltransferase 2 activity with respect to an N-glycan or O-glycan and do not display at least one activity selected from a ?-mannosyltransferase 1, 3, and 4 activity to produce recombinant proteins and glycoproteins. These recombinant Pichia pastoris strains can produce proteins and glycoproteins that lack detectable ?-mannosidase resistant ?-mannose residues thereon and thus, lack cross binding activity to antibodies against host cell antigens. Further described are methods for producing bi-sialylated human erythropoietin in Pichia pastoris that lack detectable cross binding activity to antibodies against host cell antigens.

Abstract: The present invention provides unstructured recombinant polymers (URPs) and proteins containing one or more of the URPs. The present invention also provides microproteins, toxins and other related proteinaceous entities, as well as genetic packages displaying these entities. The present invention also provides recombinant polypeptides including vectors encoding the subject proteinaceous entities, as well as host cells comprising the vectors. The subject compositions have a variety of utilities including a range of pharmaceutical applications.

Abstract: Fibrin sealant (FIBRINGLURAAS®) consisting of a kit of lyophilized high concentrate fribinogen intentionally enriched and preserved with fibronolysis inhibitor A1AT, either non-heated or heating to at least 1° C. and above, preferably at least 101° C., and lyophilized thrombin used to compound glue membrane, the diameter of which is less than 10 micrometers the actual size of the glue membrane of the fibrin sealant (FIBRINGLURAAS®) is from 0.6 ?m, to 101° C. heating 0.005 micrometers. Thrombin, a protein, contains good healthy cells. High concentrate fibrinogen, another protein, contains good healthy cells. AFOD (HDL ApoA1), another protein, contains good healthy cells and its topical applications for all solid tumor cancers.

Abstract: The invention provides a method for producing polypeptide protein products and nucleic acid products having reduced levels of antigenicity in an animal being treated with a biologic product. Somatic cells are isolated from an animal, transformed into pluripotent stem cells, transfected with a nucleic acid(s) of interest, and re-differentiated towards somatic cells known to be high level producers of the desired nucleic acid product. The invention can be used to derive a general cell line to treat populations, racial specific cell lines to treat ethnic groups, or patient specific cell lines to treat individuals. Additionally, the invention provides a method to allow induced pluripotent stem cells to be re-differentiated towards their somatic cell of origin so that the cells can be used to therapeutically treat an animal without resulting teratoma formation.

Abstract: One aspect of the invention contemplates a mutant E-WE thrombin precursor that contains the SEQ ID NO:1 amino acid residue sequence. Another aspect contemplates a thrombin precursor that contains the amino acid residue sequence Asp/Glu-Gly-Arg at positions 325, 326 and 327 based on the preprothrombin sequence. A third aspect contemplates a thrombin precursor that contains the SEQ ID NO:1 amino acid residue sequence as well as the amino acid residue sequence Asp/Glu Gly Arg at positions 325, 326 and 327 based on the preprothrombin sequence. Also contemplated is a composition that contains an effective amount of mutant thrombin dissolved or dispersed in a pharmaceutically acceptable carrier. A method is also disclosed for enhancing treating and preventing thrombosis in a mammal in need using that composition.

Abstract: The present invention relates to compositions and methods for producing recombinant human thrombin using recombinant ecarin.

Abstract: The invention provides a nucleic acid cassette comprising components in the following structure: A-B-C, wherein “A” is a nucleic acid sequence encoding a light chain of a first antibody (or antigen binding domain thereof), “B” is a nucleic acid sequence encoding a 2A peptide, “C” is a nucleic acid sequence encoding a heavy chain of a second antibody (or antigen binding domain thereof), and “-” is a phosphodiester or phosphorothioate bond. Also provided is a nucleic acid cassette with the structure A-p-B-C, where “p” is a nucleic acid encoding a protease recognition site, Also provided are methods for making recombinant antibodies using the nucleic acid cassette of the invention, cells and vector comprising the nucleic acid cassette of the invention, and kits for making the nucleic acid cassette of the invention.

Abstract: An assembly capable of capturing and purifying expressed biological products during or at the end of a bioreaction cycle is disclosed wherein a binding resin is kept separated from the contents of the bioreactor allowing capturing, harvesting and purification of biological products in a bioreactor; the invention additionally provides means of removing undesirable metabolic products as well as provides for efficient loading of chromatography columns.

Abstract: The present invention relates to matrices (e.g., fibrin-alginate matrices; fibrin-alginate-matrigel matrices) for culture of cells, organs (e.g., ovary or fragment thereof), cells and cell aggregates (e.g., ovarian follicles, embryoid bodies), and tissues. In some embodiments, protease inhibitors e.g., aprotinin) are used to prevent the degradation of fibrin.

Abstract: The present disclosure relates to a method for preparing a composition or a concentrate of a prothrombic complex that includes the II, VII, IX and X coagulation factors, including providing a supernatant of a plasma cryoprecipitate, applying the supernatant on an anion-exchange resin for producing an eluate containing the complex and proteins having a high molecular weight, and applying the eluate on a hydroxyapatite column for producing a second eluate containing the complex. The disclosure also relates to a composition that can be produced by the method.

Abstract: Methods of generating thrombin and methods of applying a clotting tissue sealant to a site on a subject are provided. A blood component comprising platelets can be obtained from the subject. A hypotonic composition is contacted with a solid matrix to form a thrombin-containing liquid, where the hypotonic composition includes water, calcium, a blood component comprising platelets, and optionally a chelator. Calcium is present in the hypotonic composition in an amount greater than the amount of calcium that can be complexed by the chelator. Thrombin-containing liquid is then separated from the hypotonic composition and can be applied to the site on the subject to form a clot, for example, by combination with fibrinogen.

Abstract: A method of alleviating pain associated with tissue damage includes applying salmon thrombin at a tissue damage site, as a single substance in liquid form, or as a powder, a foam, and/or a gel that includes salmon thrombin. A pain relief substance includes a salmon thrombin preparation.