Abstract: The embodiments described herein pertain to cells, and methods for preparing cells, that can be used as biocatalysts by altering enzymes that compete for a substrate or product of a pathway of interest such that the targeted enzyme is sensitive to a site-specific protease, which protease is expressed but relocated in the cell to a site where it is not in contact with the targeted enzyme in the intact cell. Upon cell lysis, the protease contacts the target enzyme, which is then inactivated by protease cleavage.

Abstract: The present invention provides a mutant 27 kDa NIa proteinase having reduced self-cleavage activity relative to the self-cleavage activity of its wild-type proteinase. The mutant has the same substrate cleavage activity as the wild-type proteinase but is more stable than the wild-type proteinase. The present invention also provides a method of obtaining large quantities of active 27 kDa NIa proteinase for use as a tool for purification of other proteins.

Abstract: The invention provides a single chain, polypeptide fusion protein, comprising: a non-cytotoxic protease, or a fragment thereof, which protease or protease fragment is capable of cleaving a protein of the exocytic fusion apparatus of a target cell; a Targeting Moiety that is capable of binding to a Binding Site on the target cell, which Binding Site is capable of undergoing endocytosis to be incorporated into an endocome within the target cell; a protease cleaving site at which site the fusion protein is cleavable by the protease, wherein the protease cleavage site is located between the non-cytotoxic protease or fragment thereof and the Targeting Moiety; and the translocation domain that is capable of translocating the protease or protease fragment from within an endosome, across the endosomal membrane and into the cytosol of the target cell.

Abstract: A UBP1 protease mutant and the sequence coding it, their application and products and the methods used to produce them may be used in the production of recombinant proteins, particularly on an industrial scale.

Abstract: Disclosed are disulphide-linked complexes of a soluble Tissue Factor (sTF) variant of SEQ ID NO:3 comprising the mutation G109C and a Factor VIIa variant of SEQ ID NO. 1, comprising the mutation Q64C and a mutation at position M306 that gives rise to a zymogen-like conformation in the Factor VIIa polypeptide. Said complexes may be used for the treatment of a coagulopathy.

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: The present invention relates to a method of selecting a protein variant having modified immunogenicity as compared to the parent protein comprising the steps obtaining antibody binding peptide sequences, using the sequences to localise epitope sequences on the 3-dimensional structure of parent protein, defining an epitope area including amino acids situated within 5 ? from the epitope amino acids constituting the epitope sequence, changing one or more of the amino acids defining the epitope area of the parent protein by genetical engineering mutations of a DNA sequence encoding the parent protein, introducing the mutated DNA sequence into a suitable host, culturing said host and expressing the protein variant, and evaluating the immunogenicity of the protein variant using the parent protein as reference. The invention further relates to the protein variant and use thereof, as well as to a method for producing said protein variant.

Abstract: Provided are methods for protein engineering, such as engineering proteases or kinases. The methods may utilize yeast display and/or ER sequestration of proteins or substrates. In some aspects, TEV proteases with altered substrate specificity, potency, and/or efficiency are provided.

Abstract: A method for treating a Parkinson's patient with digestive/pancreatic enzymes involves administering an effective amount of digestive/pancreatic enzymes to an individual having the disorder in order to improve a symptom of the disorder. In addition, a method is provided for determining whether an individual has, or may develop, Parkinson's disease or related dysautonomic disorders and for determining whether an individual will benefit from the administration of pancreatic/digestive enzymes to treat the dysautonomic disorder.

Abstract: The present invention relates to non-catalytic carbohydrate-binding modules (CBM) belonging to a new family of CBM's. A CBM of the invention was found attached to a glycosyl hydrolase family 61 (GH61) polypeptide and was shown to have little homology with known CBM's indicating that it is the first known member of a new family of CBM's. The present invention further relates to CBM's preferably exhibiting binding affinity for cellulose; to a method of producing such CBM's; and to methods for using such CBM's in the textile, detergent and cellulose fiber processing industries, for purification of polypeptides, immobilisation of active enzymes, baking, manufacturing of biofuel, modification of plant cell walls.

Abstract: Improved anti-HCV immunogens and nucleic acid molecules that encode them are disclosed. Immunogens disclosed include those having consensus HCV genotype 1a/1b NS3 and NS4A. Pharmaceutical composition, recombinant vaccines comprising and live attenuated vaccines are disclosed as well methods of inducing an immune response in an individual against HCV are disclosed.

Abstract: The present invention provides a synthetic gene encoding the human enterokinase light chain protein, a recombinant vector comprising the synthetic gene encoding the protein, a plant cell transformed with the recombinant vector, a method for producing the human enterokinase light chain protein in a plant by using the recombinant vector, a method for producing a plant producing the human enterokinase light chain protein by transforming a plant cell with the recombinant vector, a plant producing the human enterokinase light chain protein which is produced by the method, and a seed thereof, and a composition for large-scale production of the human enterokinase light chain protein in a plant, in which the composition comprises the synthetic gene encoding the human enterokinase light chain protein.

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: The invention provides vitamin K-dependent polypeptides with enhanced membrane binding affinity. These polypeptides can be used to modulate clot formation in mammals. Methods of modulating clot formation in mammals are also described.

Abstract: The invention relates to recombinant nucleic acid and polypeptides encoding collagenase I and collagenase II, methods for the preparation thereof and methods for the use thereof. The invention also encompasses methods related to releasing a composition comprising collagenase prior to therapeutic administration.

Abstract: This patent application discloses and describes proteins found to be differentially expressed between primary tumor breast cancer cells histologicaly defined as early stage (stage 0) breast cancer and primary breast cancer cells histologicaly defined as late stage (stage 3) breast cancer. These proteins can be used either individually or in specific combinations in diagnostic and prognostic protein assays on various biological samples from breast cancer patients to indicate the that a breast cancer patient's cancer is in an early, non-aggressive stage or in a late, aggressive stage. Determination of differential expression of these proteins can also be useful for indicating additional therapies to combat the aggressiveness of late stage breast cancer. The full length intact proteins can be assayed or peptides derived from these proteins can be assayed as reporters for these proteins.

Abstract: A use of vivapain-4 (VX-4), which is a cysteine protease of Plasmodium vivax, showing pH-dependent switching of substrate specificity, is provided. More specifically, a method of treating a parasitic disease caused by Plasmodium vivax by inhibiting VX-4; a method of screening a protease inhibitor acting on VX-4, wherein the protease inhibitor is useful as an anti-malarial agent acting on Plasmodium species, for example, Plasmodium vivax; and a method of identifying the activity of VX-4, are provided.

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: In a microbial fermentation, the aim is to increase the product yield of protein. This is achieved by a method in which an expression construct is introduced into a microorganism of the species Bacillus pumilus which comprises a promoter and a nucleic acid coding for the protein, and the protein is expressed in said expression construct.

Abstract: An enzyme useful for producing cyclic peptides from linear peptide precursors and a gene encoding the enzyme are described. The enzyme is particularly useful for producing segetalins from linear presegetalin precursors. The linear presegetalin precursors may be derived from other linear presegetalin precursors farther upstream in the biosynthetic synthesis of the segetalin.

Abstract: The aim of the invention is to improve the secretion of a protein from a host cell in order to increase the product yield of protein in a fermentation process. This is achieved by an expression vector comprising a) a promoter sequence and b) a nucleic acid sequence that codes for a protein. The protein comprises a signal peptide and an additional amino acid sequence, and the signal peptide comprises an amino acid sequence that is at least 80% identical to the amino acid sequence specified in SEQ ID NO 2, at least 80% identical to the amino acid sequence specified in SEQ ID NO 4, at least 80% identical to the amino acid sequence specified in SEQ ID NO 6, or the signal peptide comprises an amino acid sequence that is structurally homologous to at least one of said sequences.

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: An expression vector is provided. The vector includes a promoter configured to drive the expression of the transgene in the cell. The vector also includes a tag sequence encoding a tag peptide directing the protein of the expressed transgene to a pre-determined location. The vector further includes a cleavage sequence encoding a peptide that is recognizable by a protease and a marker gene configured to encoding a protein to indicate the expression of the transgene.

Abstract: The present invention provides a therapeutic agent for Alzheimer's disease. The therapeutic agent contains membrane vesicles (exosomes) of adipose tissue-derived mesenchymal stem cells, and the membrane vesicles (exosomes) contain neprilysin. Through the research by the inventors of the present invention, it was revealed that exosomes secreted by human adipose tissue-derived mesenchymal stem cells contain neprilysin, which degrades amyloid-? as a pathogenic protein of Alzheimer's disease. When exosomes secreted by human adipose tissue-derived mesenchymal stem cells were administered to the brains of Alzheimer's disease model mice, the generation of amyloid-? was inhibited.

Abstract: Compositions for regenerating tissue and wound repair, among other applications, are described.

Abstract: The invention relates to a method for synthesizing a peptide by enzymatically preparing an ester or thioester from (i) an N-terminal protected amino acid or an N-terminal protected peptide where either can have a protected C-terminal ester group and (ii) an alcohol represented by the formula HO—CX2—Z or a thiol represented by the formula HS—CX2—Z, each X independently representing a halogen atom or a hydrogen atom; and Z represents an electron withdrawing group comprising at least one sp3-hybridized carbon comprising at least two substituents comprising a heteroatom directly attached to the at least one sp3-hybridized carbon or at least one sp2-hybridized carbon comprising one or two substituents comprising a heteroatom directly attached to the at least one sp2-hybridized carbon, and enzymatically coupling the prepared ester or thioester with an optionally C-terminal protected amino acid or with an optionally C-terminal protected peptide in a medium comprising 2 wt. % water or less.

Abstract: The present invention provides improved methods and compositions for RNA isolation. In particular embodiments the present invention concerns the use of methods and compositions for the isolation of full-length RNA from fixed tissue samples. The present invention provides methods for digesting and extracting RNA from a fixed tissue sample.

Abstract: Disclosed is a method for removing endotoxin from proteins. Also disclosed are products made by using the method. The method may be used, for example, to produce endotoxin-free lactoferrin. Bovine milk-derived lactoferrin may be produced in commercial quantities by the method, and endotoxin-free bovine lactoferrin may be used for a variety of therapeutic uses, including improving wound healing.

Abstract: Screening assays and methods of using same for screening to identify modulator agents or compounds that affect matrix metalloproteinase-2 (MMP-2) mediated activation of toll-like receptor-2 (TLR-2) are described herein. Pharmaceutical and immunogenic compositions comprising agents or compounds that modulate MMP-2 mediated activation of TLR-2 are also encompassed. Methods for modulating MMP-2 mediated activation of TLR-2 using MMP-2 peptides in pharmaceutical and immunogenic compositions, as well as vaccines, are also envisioned. Melanoma is an exemplary tumor type that expresses MMP-2 and for which such pharmaceutical and immunogenic compositions, as well as vaccines, would confer benefit to patients. Also encompassed are methods for reducing MMP-2 mediated activation of TLR-2 and downstream signaling therefrom so as to achieve more effective T cell responses to MMP-2 expressing tumors.

Abstract: The invention provides methods and systems for production of recombinant protein, and particularly, for production of recombinant protein from inclusion bodies. For example, in one aspect, the method comprises providing a protein preparation comprising inclusion bodies, preparing an inclusion body dispersion, and exposing the protein preparation to high pressure in a pressure vessel, to disaggregate and refold the inclusion body protein.

Abstract: The present invention relates to isolated polypeptides having protease activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides in e.g. animal feed and detergents.

Abstract: Subtilase variants having an improved wash performance on egg stains. These subtilases are useful exhibiting excellent or improved wash performance on egg stains when used in e.g. cleaning or detergent compositions, such as laundry detergent compositions and dish wash compositions, including automatic dish wash compositions.

Abstract: Polypeptide agents useful in the treatment of IgA1 deposition diseases and methods of using such polypeptide agents. Methods of screening for inhibitors of IgA1 proteases and agents that inhibit IgA1 proteases are also disclosed.

Abstract: The specification discloses Clostridial toxins or Clostridial toxin chimeras comprising an inactivation cleavage site, polynucleotide molecules encoding such toxins or chimeras, compositions comprising such toxins or chimeras, and method of producing such toxins or chimeras.

Abstract: Reagents and methods for the digital analysis of proteins or peptides are provided. Specifically provided herein are proteins for identifying the N-terminal amino acid or N-terminal phosphorylated amino acid of a polypeptide. Also, an enzyme for use in the cleavage step of the Edman degradation reaction and a method for using this enzyme are described.

Abstract: The disclosure provides fusion proteins containing N-terminal signal peptides fused to immunogenic polypeptides. The immunogenic polypeptides may be from viruses, bacteria, or fungi. The disclosure also provides elevated expression of the immunogenic polypeptides using the N-terminal signal peptide. The N-terminal signal peptides enhance synthesis of the protein, particularly where the protein is neither a secretory nor a transmembrane peptide. The fusion proteins may be used to diagnose disease and to induce immune responses.

Abstract: The invention provides methods, compositions, systems, and kits that include an enzyme/substrate co-delivery system. The liquid delivery system includes at least one enzyme encapsulated in a water-soluble polymeric matrix and a substrate for the enzyme in a carrier liquid in which the polymeric matrix is insoluble. When water is added, the polymeric matrix is solubilized and enzyme is released from the matrix, permitting catalytic action upon the substrate.

Abstract: Improved anti-HCV immunogens and nucleic acid molecules that encode them are disclosed. Immunogens disclosed include those having consensus HCV genotype 1 a/1 b NS3 and NS4A. Pharmaceutical composition, recombinant vaccines comprising and live attenuated vaccines are disclosed as well methods of inducing an immune response in an individual against HCV are disclosed.

Abstract: Methods and reagents for the installation of click chemistry handles on target proteins are provided, as well as modified proteins comprising click chemistry handles. Further, chimeric proteins, for example, bi-specific antibodies, that comprise two proteins conjugated via click chemistry, as well as methods for their generation and use are disclosed herein.

Abstract: Provided are methods for and compounds for modulating the complement system. In particular, compounds are provided that inhibit complement activation and compounds are provided that promote complement activation. The compounds are therapeutics by virtue of their effects on the complement system. Hence, the compounds that inhibit complement activation can be used for treatment of ischemic and reperfusion disorders, including myocardial infarction and stroke, sepsis, autoimmune diseases, inflammatory diseases and diseases with an inflammatory component, including Alzheimer's Disease and other neurodegenerative disorders.

Abstract: Novel fiber processing methods and the products obtained therefrom are disclosed. Methods may include thermochemical and/or enzymatic hydrolysis of fiber feedstocks including distillers' dried grains, distillers' dried grains with solubles, soyhull, miscanthus and switchgrass. Enzymatic hydrolysis includes hydrolysis with cellulase, hemicellulase, and protease.

Abstract: The present invention provides modular extracellular sensors, nucleic acids encoding such sensors, and cells expressing such sensors, and methods of employing such sensors and cells for detecting extracellular ligands. In certain embodiments, the extracellular sensors comprise a ligand binding domain, a transmembrane domain, a protease domain, a protease cleavage site, and a transcription factor. In other embodiments, a pair of extracellular receptors is provided where both receptors contain a ligand binding domain and transmembrane domain, and one receptor contains a protease cleavage site and a transcription factor and the other receptor contains a protease domain.

Abstract: Disclosed are a method for rapid screening of suitable translational fusion partners (TFPs) capable of inducing expression or secretory production of non-producible proteins, which are difficult to produce in conventional recombinant production methods, from a variety of genetic sources, and protein secretion-inducing TFPs obtained using the method.

Abstract: The present invention consists of a two part system for controlling pet odor. The first part is an enzyme spray with a high concentration of microbes that consume ammonia. The second part of the system is absorbent granules that also contain the ammonia consuming microbes. When used in conjunction with each other, the system is ten times more effective at controlling pet odor than comparable one part systems. The microbes can penetrate into the cracks and crevices of most surfaces to further reduce the source of pet odors. The system also works to control moisture which allows for proper pet hygiene by reducing molds, fungus, insects, and parasite infestations. The system is a non-toxic and environmentally friendly product that will not harm pets if ingested. The system can be used with cat litter boxes, pet bedding, dog kennels, or any indoor area where pets expel their waste.

Abstract: Methods for extracting high quality nucleic acids from a heterogenous collection of nucleic acid-containing materials from a biological sample are disclosed. The heterogenous collection of nucleic-acid containing materials may contain cells or microvesicles, or both. The extractions obtained by the methods described herein are characterized by high yield and high integrity, making the extracted nucleic acids useful for various applications in which high quality nucleic acid extractions are preferred, e.g., a diagnosis, prognosis, or therapy evaluation for a medical condition.

Abstract: The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

Abstract: The present invention relates to isolated polypeptides having endoglucanase activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: The present invention relates to isolated polypeptides having cellobiohydrolase activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: The present invention relates to isolated polypeptides having cellobiohydrolase activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: A combination enzyme product consisting of a glutamine specific endoprotease and a prolyl endopeptidase is provided. Both enzymes are active and stable in the stomach and can therefore be administered as lyophilized powders or simple capsules/tablets. A ratio of the two enzymes is used to maximize their synergy.