Abstract: The present invention relates to protease variants and methods for obtaining protease variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Abstract: The present invention relates to protease variants and methods for obtaining protease variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Abstract: This disclosure relates to methods for isolating antibodies from cell-free culture supernatant.

Abstract: The present invention relates to protease variants and methods for obtaining protease variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Abstract: The present invention relates to detergent compositions comprising protease variants and methods for obtaining such detergent compositions. The present invention also relates to the use of such detergent compositions, especially in laundry or in hard surface cleaning applications.

Abstract: The present invention relates to protease variants and methods for obtaining protease variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Abstract: New thrombolytic protein molecules such as recombinant staphylokinase or streptokinase, urokinase, tissue plasminogen activator and the like, and suitable variants thereof, for targeting to brain tissue or any other tissue by either fusing to, or by synthesizing the candidate thrombolytic molecule(s) with a protein sequence comprising a strong amphipathic alpha helix containing protein transduction domain. Thrombolytic protein molecule(s) so engineered with the protein transduction domain is useful for enhanced uptake of such protein thrombolytic molecule(s) across the cell membranes and tissues including the blood brain barrier and find their use in the treatment of vascular thrombosis including cerebrovascular disorders caused by cerebral thrombosis or cerebral haemorrhage when used a as a therapeutic. The design and processes for cloning, expression, purification and protein transduction of such proteins across cell membranes.

Abstract: The disclosure relates to a Gram negative bacterial cell that is transformed with a nucleic acid molecule that encodes a Gram positive twin-arginine translocase and including methods for the production of polypeptides.

Abstract: This disclosure relates to a method of generating conditionally active biologic proteins from wild type proteins, in particular therapeutic proteins, which are reversibly or irreversibly inactivated at the wild type normal physiological conditions. For example, evolved proteins are virtually inactive at body temperature, but are active at lower temperatures.

Abstract: The present invention demonstrates the utility of carbonic acid amides such as urea or its derivatives, carbamates, carbodiimides & thiocarbamides as nitrogenous supplements in fermentation media for production of recombinant proteins to achieve enhanced bioconversion rates and peptides like insulin and insulin analogues, exendin and enzymes such as lipase using methanol inducible fungal expression systems such as Pichia.

Abstract: The present invention discloses novel hybrid proteins that have both plasminogen activator and anti-thrombotic properties, including clot specific action, that renders these as highly advantageous for the treatment of circulatory disorders involving fibrin clot formation due to underlying tissue damage in the blood vessels leading to myocardial infarction, strokes etc. Also disclosed are new proteins, and methods of obtaining the same, that help to dissolve blood clots by activating plasminogen in a plasmin or thrombin dependent manner and also inhibit both the activity and generation of thrombin through the intrinsic pathway of blood coagulation.

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 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: The present invention relates to novel mutants of Streptokinase, its functional fragments and covalently modified forms. Methods are provided for the preparation of the bacterial plasminogen activator protein, Streptokinase its muteins, species variants and their covalently modified variants that are characterized by improved therapeutic properties, such as increased proteolytic stability, extended plasma half-lives, reduced immuno-reactivity and enhanced fibrin clot specificity. The method involves either incorporating additional cysteine residues, or substituting cysteine residues for naturally occurring amino acids into non-essential regions of the protein such that the catalytic activity of the resultant protein remains largely unaltered. These cysteine variants were further modified by covalently attaching a cysteine reactive polymer such as polyethylene glycol (PEG) or sulfhydryl-reactive moieties from a group that includes fluorophore, spin labels or other small conjugates.

Abstract: This disclosure relates to a method of generating conditionally active biologic proteins from wild type proteins, in particular therapeutic proteins, which are reversibly or irreversibly inactivated at the wild type normal physiological conditions. For example, evolved proteins are virtually inactive at body temperature, but are active at lower temperatures.

Abstract: The present invention relates to the use of G-CSF and derivatives thereof for extending the therapeutic window of subsequent thrombolytic treatment of acute stroke, and thereby, allowing the diagnostic examinations which are necessary prior to the thrombolytic treatment in order to avoid hemorrhagic and other severe adverse side effects of the thrombolysis.

Abstract: The present invention relates to novel mutants of Streptokinase, its functional fragments and covalently modified forms. Methods are provided for the preparation of the bacterial plasminogen activator protein, Streptokinase its muteins, species variants and their covalently modified variants that are characterized by improved therapeutic properties, such as increased proteolytic stability, extended plasma half-lives, reduced immuno-reactivity and enhanced fibrin clot specificity. The method involves either incorporating additional cysteine residues, or substituting cysteine residues for naturally occurring amino acids into non-essential regions of the protein such that the catalytic activity of the resultant protein remains largely unaltered. These cysteine variants were further modified by covalently attaching a cysteine reactive polymer such as polyethylene glycol (PEG) or sulfhydryl-reactive moieties from a group that includes fluorophore, spin labels or other small conjugates.

Abstract: The present invention demonstrates the utility of carbonic acid amides such as urea or its derivatives, carbamates, carbodiimides & thiocarbamides as nitrogenous supplements in fermentation media for production of recombinant proteins to achieve enhanced bioconversion rates and peptides like insulin and insulin analogues, exendin and enzymes such as lipase using methanol inducible fungal expression systems such as Pichia.

Abstract: The invention provides compositions and methods of identifying, modifying and producing modified target molecules, including therapeutic molecules by modification with non-natural amino acids. Certain aspects of the invention include methods of adding a chemical moiety to a target molecule, and the compositions resulting therefrom. Certain aspects of the invention also relate to kits for identifying, modifying and producing modified target molecules described herein.

Abstract: The present invention relates to novel mutants of Streptokinase, its functional fragments and covalently modified forms. Methods are provided for the preparation of the bacterial plasminogen activator protein, Streptokinase its muteins, species variants and their covalently modified variants that are characterized by improved therapeutic properties, such as increased proteolytic stability, extended plasma half-lives, reduced immuno-reactivity and enhanced fibrin clot specificity. The method involves either incorporating additional cysteine residues, or substituting cysteine residues for naturally occurring amino acids into non-essential regions of the protein such that the catalytic activity of the resultant protein remains largely unaltered. These cysteine variants were further modified by covalently attaching a cysteine reactive polymer such as polyethylene glycol (PEG) or sulfhydryl-reactive moieties from a group that includes fluorophore, spin labels or other small conjugates.

Abstract: New thrombolytic protein molecules such as recombinant staphylokinase or streptokinase, urokinase, tissue plasminogen activator and the like, and suitable variants thereof, for targeting to brain tissue or any other tissue by either fusing to, or by synthesizing the candidate thrombolytic molecule(s) with a protein sequence comprising a strong amphipathic alpha helix containing protein transduction domain. Thrombolytic protein molecule(s) so engineered with the protein transduction domain is useful for enhanced uptake of such protein thrombolytic molecule(s) across the cell membranes and tissues including the blood brain barrier and find their use in the treatment of vascular thrombosis including cerebrovascular disorders caused by cerebral thrombosis or cerebral haemorrhage when used a as a therapeutic. The design and processes for cloning, expression, purification and protein transduction of such proteins across cell membranes.

Abstract: An improved process for the production of streptokinase using a genetically engineered strain of Escherichia coli which overproduces streptokinase intracellularly and more particularly, the overall process disclosed herein, concerns with an improvement in the fermentative production of streptokinase using an optimized growth medium mainly comprised of simple salts and trace-elements; thus, in principal, the present process constitutes an improved and more economical means for the production of streptokinase which may be useful in thrombolytic therapy.

Abstract: Disclosed is both a process for producing a reversibly inactive acidified plasmin by activating plasminogen and a process for producing a purified plasminogen. The produced plasmin is isolated and stored with a low pH-buffering capacity agent to provide a substantially stable formulation. The purified plasminogen is typically purified from a fraction obtained in the separation of immunoglobulin from Fraction II+III chromatographic process and eluted at a low pH. The reversibly inactive acidified plasmin may be used in the administration of a thrombolytic therapy.

Abstract: The present invention relates to a nucleotide sequence of expression cassette OXY-1 of SEQ ID No. 1, a modified staphylokinase SAK-2 gene of SEQ ID No. 2, a peptide sequence of modified staphylokinase SAK-2 gene, of SEQ ID No. 3, three plasmids having International Deposition Nos. BPL-0019, BPL-0020, and BPL-0021, and their corresponding three recombinant E. Coli; also invention relates to a process for over-producing staphylokinase and its analogues by modulating level of oxygen of its growth medium in a host system, and lastly, a method of dissolving blood clot in a subject in need thereof.

Abstract: The disclosure provides fusion polypeptides and constructs useful in targeting molecules including diagnostics and therapeutics to a cell type of interest. The fusion constructs include a protein transduction domain, a ligand domain and a cargo domain. Also provided are methods of treating disease and disorders such as cell proliferative disorders.

Abstract: A target protein is rendered less immunogenic to a given species by (a) determining at least part of the amino acid sequence of the target protein; (b) identifying in the amino acid sequence one or more potential epitopes for T-cells (“T-cell epitopes”) of the given species; and (c) modifying the amino acid sequence to eliminate at least one of the T-cell epitopes identified in step (b) to reduce the immunogenicity of the protein when exposed to the immune system of the given species.

Abstract: A reagent for the detection of an extracellular enzymatically active protein produced by a beta-hemolytic streptococcus bacteria found in a host biological fluid includes a proteinaceous substrate or a cholesterol-containing membrane substrate for the extracellular protein. The substrate is nonspecific within the groups of beta-hemolytic streptococcus bacterium and is in contact with an inert solid matrix. Upon reaction between the streptococcus enzymatically activate protein and the substrate, a color change discernable by an unaided human eye results. Extracellular streptococcus protein found in saliva represents a less invasive source of biological fluid for the determination as to whether a host suffers acute pharyngitis.

Abstract: The present invention relates to methods for producing variants of a parent TY145 subtilase and of a parent BPN? subtilase and to TY145 and BPN? variants having altered properties as compared to the parent TY145/BPN? subtilase.

Abstract: The present invention provides polynucleotides encoding clot-specific streptokinase proteins possessing altered plasminogen characteristics, including enhanced fibrin selectivity. The kinetics of plasminogen activation by these proteins are distinct from those of natural streptokinase, in that there is a temporary delay or lag in the initial rate of catalytic conversion of plasminogen to plasmin. Also disclosed are processes for preparing the proteins.

Abstract: An improved process for the production of streptokinase using a genetically engineered strain of Escherichia coli which overproduces streptokinase intracellularly and more particularly, the overall process disclosed herein, concerns with an improvement in the fermentative production of streptokinase using an optimized growth medium mainly comprised of simple salts and trace-elements; thus, in principal, the present process constitutes an improved and more economical means for the production of streptokinase which may be useful in thrombolytic therapy.

Abstract: A process for inhibiting vascular proliferation separately introduces components into the eye to generate plasmin in the eye in amounts to induce complete posterior vitreous detachment where the vitreoretinal interface is devoid of cortical vitreous remnants. The process administers a combination of lysine-plasminogen, at least one recombinant plasminogen activator selected from the group consisting of urokinase, streptokinase, tissue plasminogen activator, chondroitinase, pro-urokinase, retavase, metaloproteinase, and thermolysin and a gaseous adjuvant to form a cavity in the vitreous. The composition is introduced into the vitreous in an amount effective to induce crosslinking of the vitreous and to induce substantially complete posterior vitreous detachment from the retina without causing inflammation of the retina.

Abstract: The present invention provides a process for economically producing N-acetylneuraminic acid without using expensive materials such as pyruvic acid and phosphoenolpyruvic acid. The process comprises: allowing (i) a culture of a microorganism having N-acetylneuraminic acid aldolase activity or N-acetylneuraminic acid synthetase activity, or a treated matter of the culture, (ii) a culture of a microorganism capable of producing pyruvic acid or a treated matter of the culture, or a culture of a microorganism capable of producing phosphoenolpyruvic acid or a treated matter of the culture, (iii) N-acetylmannosamine, and (iv) an energy source which is necessary for the formation of pyruvic acid or phosphoenolpyruvic acid to be present in an aqueous medium to form and accumulate N-acetylneuraminic acid in the aqueous medium; and recovering N-acetylneuraminic acid from the aqueous medium.

Abstract: The present invention relates to a novel microorganism Streptomyces megasporus SD5. The present invention also relates to a process for the isolation of said Streptomyces megasporus SD5. The invention also relates to a novel fibrinolytic enzyme actinokinase extracted from said microorganism and to a process for the extraction of said enzyme. In another aspect, the invention also pertains to a method for the treatment of thrombolytic disorders using said enzyme.

Abstract: A process for inhibiting vascular proliferation introduces a composition into the eye inducing posterior vitreous detachment. The composition includes a combination of plasminogen, a collagen crosslinking agent and at least one plasminogen activator selected from the group consisting of urokinase, streptokinase, tissue plasminogen activator, chondroitinase, pro-urokinase, retavase, metaloproteinase, and thermolysin. The composition is introduced into the vitreous in an amount effective to induce crosslinking of the vitreous and to induce substantially complete or partial posterior vitreous detachment from the retina without causing inflammation of the retina. In one embodiment, a gaseous material is introduced into the vitreous before or simultaneously with the composition to compress the vitreous against the retina while the composition induces the posterior vitreous detachment.

Abstract: Compositions and methods for prevention and treatment of uncontrolled formation of intravascular fibrin clots are provided wherein fibrinolytic or anticoagulant drugs are biocompatibly coupled to red blood cell carriers.

Abstract: The present invention is related to the field of biotechnology and genetic engineering techniques, particularly to a method for obtaining mutants obtain from streptokinase, to the molecules obtained from this method, as well as the expression vectors and microorganisms for recombinant obtaining. The object of the present invention is to achieve streptokinase mutants from modifications of skc-2 gene coding for streptokinase SKC-2 (Heberkinase®), such that the obtained mutants conserve their capacity for plasminogen activator complex formation having reduced antigenicity, that could constitute preferred alternatives to native streptokinase for thrombolytic therapy. The molecules obtained from present invention can be used in the treatment of disorders as myocardial infarct, pulmonary thromboembolism, surgical complications and other cases of thrombosis.

Abstract: A pharmaceutical preparation contains protein C and a thrombolytically active substance that does not activate protein C. This preparation prevents reocclusion usually occurring in the course of thrombolysis therapy.

Abstract: Novel vaccines for use against &bgr;-hemolytic Streptococcus colonization or infection are disclosed. The vaccines contain an immunogenic amount of a variant of strepococcal C5a peptidase (SCP). Also disclosed is a method of protecting a susceptible mammal against &bgr;-hemolytic Streptococcus colonization or infection by administering such a vaccine. Enzymatically inactive SCP, and polynucleotides encoding these SCP proteins are further disclosed.

Abstract: The present invention is related to the field of biotechnology and genetic engineering techniques, particularly to a method for obtaining mutants obtain from streptokinase, to the molecules obtained from this method, as well as the expression vectors and microorganisms for recombinant obtaining. The object of the present invention is to achieve streptokinase mutants from modifications of skc-2 gene coding for streptokinase SKC-2 (Heberkinase®), such that the obtained mutants conserve their capacity for plasminogen activator complex formation having reduced antigenicity, that could constitute preferred alternatives to native streptokinase for thrombolytic therapy. The molecules obtained from present invention can be used in the treatment of disorders as myocardial infarct, pulmonary thromboembolism, surgical complications and other cases of thrombosis.

Abstract: The present invention provides methods for diagnosing and/or monitoring thrombophilic disease in a patient that can result from the antiphospholipid antibody syndrome (aPL syndrome). The methods of the invention are premised on the inhibition of binding of an anticoagulant protein, annexin, preferably annexin-V, to phospholipids by antiphospholipid (aPL) antibodies in a patient blood sample.

Abstract: The present invention relates to a plasminogen activator inhibitor, an external preparation for skin comprising the same, a method for improving rough skin and in particular, to improvement of an effective inorganic component.

Abstract: The invention relates to novel protease-inhibitors which are obtainable from leeches. It also relates to uses thereof, for instance as a medicament, thus pharmaceutical preparations are provided, as are derivatives, mutants, genes encoding, vectors comprising and cells provided with such genes and/or vectors. In particular the invention relates to a family of proteinaceous protease-inhibitors having a molecular weight of about 5.5 kD and the following primary sequences: DDNCGGKVCSKGQLCHDGHCECTPIRCLIFCPNGFAVDENGCELPCSCKHQ, DDDCGGQVCSKGQLCVDGQCKCTPIRCRIYCPKGFEVDENGCELPCTCLQ and DGNCGGQVCSKGQLCVDGQCKCTPIRCRIYCPKGFEVDENGCELPCTCLQ. This invention also relates to HIV-inhibitors and other therapeutically interesting, low molecular weight, and low antigenic substances from leeches.

Abstract: A pharmaceutical composition in a preferred embodiment comprises an isolated bacterial protein that induces fibrin-dependent plasminogen activation, and methods for dissolving blood clots in a subject use such a composition. Embodiments also include a nucleic acid encoding such a bacterial protein, a nucleic acid encoding such a bacterial protein as a fusion to another protein, an expression vector with the nucleic acid, and a host cell transformed with the expression vector.

Abstract: The present invention describes a novel polypeptide, and methods of its use in effective thrombolytic therapy in the treatment of coronary and pulmonary thrombosis. Its use is also disclosed in vaccines to abrogate a streptococcal infection. Pharmaceutical compositions containing the novel polypeptide are included. One particular form of the novel polypeptide is streptococcal surface enolase (SEN), a specific binding protein for human plasmin and/or human plasminogen on group A streptococci that displays classical &agr;-enolase activity, i.e., it can catalyze the dehydration of D-glycerate-2-phosphate to phosphoenolpyruvate. In addition, SEN impedes the inhibition of the fibrinolytic activity of plasmin by &agr;2-antiplasmin and can bind plasminogen without preventing streptokinase from cleaving this plasmin precursor.

Abstract: Streptokinase derivatives having platelet glycoprotein binding domains adjacent to the termini of the streptokinase sequence. These derivatives produce higher local concentrations of plasmin in vivo as compared to unmodified streptokinase. Certain of the derivatives have high affinity for the GPIIB/IIIA receptor and low affinity for the fibronectin and vitronectin receptors. Others have substantially equivalent affinity for all three receptors. The derivatives are useful in treating thromboembolic disorders. The streptokinase derivatives can be made by recombinant techniques or by chemical synthesis or conjugation.

Abstract: Platelet-specific, chimeric immunoglobulin and immunoglobulin fragments are described. The chimeric molecules are made up of a nonhuman antigen binding region and a human constant region. Preferred immunoglobulins are specific for glycoprotein IIb/IIIa receptor in its complexed form; they block ligand binding to the receptor and prevent platelet aggregation. The immunoglobulins are useful in anti-thrombotic therapy when administered alone or in conjunction with thrombolytic agents, as well as in thrombus imaging.

Abstract: The invention relates to the use of thromboembolic proteins. These contain, as their only structural portion effecting thrombolytic activity, the protease domain of human tissue type plasminogen activator. These derivatives show reduced side effects, such as a reduction in bleeding while demonstrating remarkable in vivo efficacy. The effect is surprising, given their in vitro properties.

Abstract: Mutants of streptokinase in which one or more amino acid residues in the region of Pro58-Lys59-Ser60-Lys61 were replaced by other amino acids become better fibrinolytic agents. The mutants are resistant to the hydrolytic inactivation by plasmin and are more efficient in inducing fibrinolysis of plasma clots.

Abstract: The present invention provides a human serine protease precursor (HSPP) and polynucleotides which encode HSPP. The invention also provides expression vectors, host cells, agonists, antisense molecules, antibodies, or antagonists. The invention also provides methods for treating disorders associated with expression of HSPP.

Abstract: The invention features modified streptokinase (SK) molecules which are resistant to plasmin cleavage including a recombinant fusion protein in which the amino terminus of SK was blocked with a peptide, a recombinant fusion protein in which an amino-terminal deleted SK was blocked with a peptide, and a mutated SK in which plasmin-cleavage sites were altered to render those sites resistant to enzymatic cleavage.