Abstract: The present invention relates to methods for pretreating biological samples for extraction of nucleic acid therefrom. The present invention employs a combination of at least one protein denaturant with one or more of the following elements to form a reaction mixture for extraction of nucleic acid: (1) at least one aprotic solvent, (2) stepwise heating, and (3) sample dilution.

Abstract: Recombinant activated protein C (APC) and APC variants with reduced anticoagulant activity were used to reduce mortality in murine models of sepsis. These models included endotoxemia and bacteremia models. We discovered that single or multiple bolus doses of APC, especially of APC variants such as RR230/231AA-APC, KKK192-194AAA-APC and 5A-APC (containing the combination of mutations present in the first two APC variants) given as a single bolus reduces 7-day mortality of mice given lethal doses of endotoxin. Administrations of a single bolus of 5A-APC after the initiation of sepsis also reduces mortality caused by LPS. 5A-APC with ?8% of normal anticoagulant activity (which has reduced risk of bleeding) reduces mortality when given as two bolus administrations at 3 hours and then at 10 hours after initiation of bacterial infection, i.e. after onset of sepsis.

Abstract: The invention relates to an ex vivo animal or challenge model as a method to identify protective (recombinant) proteins and rapidly measure protective immunity in intestinal segments directed against parasites and vaccines directed against parasitic infections. The invention further relates to vaccines directed against infection with parasites, such as Fasciola hepatica, which vaccines contain protective (recombinant) proteins identified and shown to be protective in studies using the ex vivo model. The invention further relates to protective (recombinant) proteins obtained from newly excysted juveniles (NEJ) of Fasciola hepatica. The protective (recombinant) protein corresponding to an NEJ protein has an apparent molecular weight of 32 kD and an N-terminal amino acid sequence comprising the sequence XXDVSWPFWDRMYNY (SEQ ID NO:1).

Abstract: A Factor X (hereinafter referred to as “FX”) with a high activity is provided. The present invention relates to a method for efficiently preparing a recombinant, two-chain FX which comprises intervening glycosylation at such an amino acid sequence that is essential for glycosylation in FX to thereby allow for expression of a recombinant FX with no glycosylation, and the recombinant FX with no glycosylation obtained by said method.

Abstract: Disclosed herein are diagnostic markers for lung cancer, isolated from serum glycoproteins. The disclosed diagnostic markers for lung cancer are specifically expressed only in the sera of lung cancer patients at high levels, and thus will be very useful for diagnosing lung cancer and estimating disease progression and treatment.

Abstract: The invention relates to the discovery of novel soluble neutral active Hyaluronidase Glycoproteins (sHASEGPs), methods of manufacture, and their use to facilitate administration of other molecules or to alleviate glycosaminoglycan associated pathologies. Minimally active polypeptide domains of the soluble, neutral active sHASEGP domains are described that include asparagine-linked sugar moieties required for a functional neutral active hyaluronidase domain. Included are modified amino-terminal leader peptides that enhance secretion of sHASEGP. The invention further comprises sialated and pegylated form of a recombinant sHASEGP to enhance stability and serum pharmacokinetics over naturally occurring slaughterhouse enzymes. Further described are suitable formulations of a substantially purified recombinant sHASEGP glycoprotein derived from a eukaryotic cell that generate the proper glycosylation required for its optimal activity.

Abstract: Described herein are methods for the solubilization and proteolytic cleavage of fusion protein aggregates, including autocatalytic fusion proteins, at pressures greater than atmospheric pressure to yield soluble target polypeptides.

Abstract: The invention provides compositions and methods for identifying and/or quantifying glycopolypeptides from human serum or plasma. The compositions and methods include a plurality of standard peptides containing glycosylation sites determined for human serum/plasma proteins.

Abstract: The present invention is generally directed to biological assays. More specifically it is directed to FRET-based assays using particularly effecting FRET pairs, methods for performing such assays and the molecules utilized in the assays. In a composition aspect, the present invention is directed to a FRET-based protease substrate selected from the list of substrates shown in the Detailed Description section above. In a method aspect, the present invention is directed to a method of performing a FRET-based assay, where the assay includes the following steps: adding a solution or suspension containing one or more different proteins to a reaction vessel; adding a liquid comprising a FRET-substrate to the reaction vessel, wherein the FRET-substrate is selected from the list of substrates shown in the Detailed Description section above. In a kit aspect, the present invention is directed to a kit for performing a FRET-based assay, wherein the kit comprises a protease substrate.

Abstract: The present invention relates to a protease having algicidal activity, a gene encoding the same and algicidal formulation comprising the same. The protease according to the present invention showed high algicidal activity. Therefore, the protease can be used treatment of red tide in marine region of red tide occurrence.

Abstract: The present invention pertains in general to Bromelain and particularly to the different active compounds contained in this complex mixture of proteins. The present invention provides recombinant expressed cysteine proteases, which are found in Bromelain. It has been found that the method for expression of the recombinant proteins is superior to the purification from Bromelain itself.

Abstract: The present invention relates to a method, a special designed cuvette (1) and an apparatus for the monitoring of spatial fibrin clot formation or dissolution in multiple samples.

Abstract: Isolated and/or recombinant enzymes that include surface binding domains, surfaces with active enzymes bound to them and methods of coupling enzymes to surfaces are provided. Enzymes can include large and/or multiple surface coupling domains for surface coupling.

Abstract: The present invention relates antidotes to anticoagulants targeting factor Xa. The antidotes are factor X and factor Xa protein derivatives that bind to the factor Xa inhibitors thereby substantially neutralizing them but do not assemble into the prothrombinase complex. The derivatives describe herein lack or have reduced intrinsic coagulant activity. Disclosed herein are methods of reversing anticoagulation, stopping or preventing bleeding in a patient that is currently undergoing anticoagulant therapy with a factor Xa inhibitor.

Abstract: The present invention relates to the crystal structure of the serine protease kallikrein 7 and to the use of this crystal structure in drug discovery. The present invention also relates to compounds binding specifically to this active site of kallikrein 7.

Abstract: The invention relates to bacteria that have increased levels of protein secretion due to genetic modification, to nucleotide sequences and gene structures containing at least one gene coding for a SecA protein having increased levels of protein secretion, to a SecA having increased levels of protein secretion, and to a method for producing desired proteins using the inventive bacteria. The invention also relates to nucleic acids coding for a SecA protein having increased levels of protein secretion, containing a SecA gene sequence or allele, a SecA homologue or derivative, or nucleotide sequences hybridising therewith and comprising at least one mutation. Surprisingly, just one mutation in a nucleotide of a SecA gene leads to increased levels of protein secretion or to protein secretion for the first time.

Abstract: 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 nociceptive sensory afferent; a Targeting Moiety that is capable of binding to a Binding Site on the nociceptive sensory afferent, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the nociceptive sensory afferent; a protease cleavage site at which site the fusion protein is cleavable by a protease, wherein the protease cleavage site is located between the non-cytotoxic protease or fragment thereof and the Targeting Moiety; and a 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 nociceptive sensory afferent. Nucleic acid sequences encoding the polypeptide fusion proteins, methods of preparing same and uses thereof are also described.

Abstract: The present invention provides a blood coagulation promoter capable of exerting both excellent blood coagulation promoting ability and excellent blood clot detaching ability, and a blood collection tube accommodating the blood coagulation promoter. The present invention provides a blood coagulation promoter which includes a hardly water soluble polyoxyalkylene derivative; a partially saponificated polyvinyl alcohol; at least one substance selected from the group consisting of adsorptive inorganic substances and hydrolases capable of hydrolyzing a bond between Arg and an arbitrary amino acid reside and/or a bond between Lys and an arbitrary amino acid residue in a peptide chain; a polyvinylpyrrolidone; and preferably a water-soluble silicone oil, and a blood collection tube in which the blood coagulation promoter is accommodated in a tubular vessel having a bottom.

Abstract: An arterial thrombosis risk factor comprising one or more of the identified mutants of coagulation factor VII activating protease (FSAP) is described. In addition, diagnostic determination methods for detecting these mutants which are identified as risk factors are described.

Abstract: The present invention is directed to novel food products, e.g., nutritional food products and infant formula, which contain one ore more enzymes selected from lipase, protease, and amylase that have been formulated/stabilized to have sustained stability in an aqueous medium. Such formulations are intended to provide a greater degree of compliance based on their ability to be incorporated into aqueous media while avoiding unstable breakdown of the enzyme and large overdosing due to expected breakdown when exposed to an aqueous environment, including saliva. Further described in the invention are additives packaged with instructions for combination with an aqueous medium, and instructions for the administration of the resulting mixture to a subject. In certain embodiments, enzyme insufficient patients, e.g., infants and elderly persons, would find particular benefit from the food products described herein.

Abstract: The present invention provides methods for engineering proteins to optimize their performance under certain environmental conditions of interest. In some embodiments, the present invention provides methods for engineering enzymes to optimize their catalytic activity under particular environmental conditions. In some preferred embodiments, the present invention provides methods for engineering enzymes to optimize their catalytic activity and/or stability under adverse environmental conditions. In some preferred embodiments, the present invention provides methods for engineering enzymes to optimize their storage stability, particularly under adverse environmental conditions. In some preferred embodiments, the present invention provides methods for altering the net surface charge and/or surface charge distribution of enzymes (e.g., metalloproteases) to obtain enzyme variants that demonstrate improved performance and/or stability in detergent formulations as compared to the starting or parent enzyme.

Abstract: Systems and methods are provided for producing a protein of interest that is typically not amenable to expression in soluble form in in vitro expression systems. In some aspects, the invention provides methods of synthesizing proteins using in vitro protein synthesis systems that include a scaffold protein such as apolipoprotein or an amphipathic alpha helix containing (“AAHC”) protein, in which higher yields of soluble protein are produced than in the absence of the scaffold protein. The scaffold proteins may be provided in an in vitro protein synthesis system associated with lipid or not associated with lipid. The scaffold protein may be provided as a protein per se or may be encoded by a nucleic acid template and co-expressed with the protein of interest. The invention also provides compositions and kits for synthesis of proteins in soluble form, in which the compositions and kits include cell extracts for protein expression and isolation.

Abstract: Disclosed herein are a materials such as a coating, an elastomer, an adhesive, a sealant, a textile finish, a wax, and a filler for such a material, wherein the material includes an enzyme such as an esterase (e.g., a lipolytic enzyme, a sulfuric ester hydrolase, an organophosphorus compound degradation enzyme), an enzyme that degrades a cell wall and/or a cell membrane component (e.g., a lysozyme, a lytic transgrycosylase, a peptidase), and/or a biocidal or biostatic peptide. Also disclosed herein are methods of decontaminating a surface comprising such a material from a chemical substrate of an enzyme such as a lipid or an organophosphorus compound, as well as reducing the growth of a microorganism on or within such a material.

Abstract: Methods for preparing Factor X, activated Factor X, inactivated factor X and inactivated factor Xa, compositions comprising Factor X and Factor Xa, inactivated Factor X and inactivated Factor Xa and methods of medical treatment using Factor X, Factor Xa, activated Factor X and inactivated Factor Xa are disclosed. The preparation methods comprise a chromatography step using an immobilised metal ion affinity chromatography substrate.

Abstract: The present invention relates to agent compositions, in particular washing and cleaning agents, that contain a protease from a bacterial strain of the species Stenotrophomonas maltophilia, and to said proteases themselves. The invention further relates to cleaning methods in which said agents are used, and to uses of said agents. The invention further relates to the manufacture and use of these proteases themselves.

Abstract: Apparatus suitable for use in the prevention and treatment of burn induced compartment syndrome facilitates the rapid and efficient deployment of enzymatic compositions to the affected limb or limbs of a burn victim suffering from a circumferential burn. The apparatus includes a flexible, water-impermeable generally cylindrical covering layer. The covering layer has an inner surface facing the limb and an outer surface. The covering layer has a proximal sealable open end through which the burned limb is inserted and a distal closed end. An enzymatic escharotomy agent is contained within the covering layer. The covering layer retains the enzymatic escharotomy agent in close proximity to the burned limb, and the apparatus is enclosed within a sterile sealed package evacuated from moisture.

Abstract: In this application is described isolated, recombinant CapD and recombinant PghP for use in digesting capsule comprising polyglutamate polymers and for treatment of infections caused by bacilli having a polyglutamate capsule, such as anthrax.

Abstract: The present invention relates to the development of new derivatives of a bacterial plasminogen activator, Staphylokinase (SAK), having one or more amino acid residues with single or multiple cysteines at the amino and/or carboxy terminal ends and their conjugation with PEG (Polyethylene Glycol), resulting in new Staphylokinase derivatives that display altered oligomeric states, enhanced thermal and protease stability and extended plasma half-life. Also included is the cloning and expression in a suitable bacterial host; purification of Staphylokinase derivatives to homogeneity and their chemical modification by integrating a PEG molecule to create new biologically active Staphylokinases having higher protein stability and improved in vivo plasma half life, that may enhance the clinical potential of Staphylokinase in thrombolytic therapy for the treatment of cardiovascular diseases.

Abstract: Provided are methods to identify modulators and in particular inhibitors of body malodour formation employing peptidase enzymes, the peptidase enzymes and corresponding nucleotide sequences, expression vectors, transfected host cells, methods of forming the peptidase enzymes and methods to prevent body malodour.

Abstract: The present invention generally relates to various nucleases and uses thereof, and in some cases, to the UPF0054 protein superfamily. Members of the UPF0054 protein superfamily, such as the E. coli protein YbeY, may possess RNase activity and may be involved in certain important cellular processes. Disruption of YbeY activity can lead to increased sensitivity to antibiotics. Accordingly, certain embodiments of the invention are directed to systems and methods for screening target compounds for activity against UPF0054 superfamily proteins. In some embodiments, the screening method allows to target compositions to be determined that show selective activity against UPF0054 superfamily proteins. Other embodiments of the invention provide for nucleases capable of site-specific cleavage of nucleic acids.

Abstract: The invention provides isolated nucleic acids molecules and proteins, designated 27411, 23413, 22438, 23553, 25278, 26212, NARC SC1, NARC 10A, NARC 1, NARC 12, NARC 13, NARC17, NARC 25, NARC 3, NARC 4, NARC 7, NARC 8, NARC 11, NARC 14A, NARC 15, NARC 16, NARC 19, NARC 20, NARC 26, NARC 27, NARC 28, NARC 30, NARC 5, NARC 6, NARC 9, NARC 10C, NARC 8B, NARC 9, NARC2A, NARC 16B, NARC 1C, NARC 1A, NARC 25, 86604 and 32222 nucleic acid molecules and proteins. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing said nucleic acid molecules, host cells into which the expression vectors have been introduced, nonhuman transgenic animals in which a said genes have been introduced or disrupted, fusion proteins, antigenic peptides and antibodies to said proteins. Diagnostic and therapeutic methods utilizing compositions of the invention are also provided.

Abstract: Disclosed is an agent for digesting a protein highly resistant to denaturation and degradation, comprising as an active ingredient an enzyme exhibiting an activity of digesting a protein highly resistant to denaturation and degradation and having the following properties: (a) activity and substrate specificity: hydrolyzing a peptide bond of a protein highly resistant to denaturation and degradation; (b) molecular weight: 31,000 (determined by SDS-polyacrylamide gel electrophoresis using a homogeneous gel having a gel concentration of 12%); (c) isoelectric point: pI 9.3 (determined by polyacrylamide gel isoelectric focusing electrophoresis); (d) optimum pH: pH 9.0 to 10.0; and (e) optimum temperature for activity: 60 to 70° C.

Abstract: A mutated severe acute respiratory syndrome-associated coronavirus 3C-like protease and use thereof for cleaving a protein that includes a cleavage site recognizable by the mutated protease to yield a polypeptide fragment of interest.

Abstract: The invention relates to bacteria that have increased levels of protein secretion due to genetic modification, to nucleotide sequences and gene structures containing at least one gene coding for a SecA protein having increased levels of protein secretion, to a SecA having increased levels of protein secretion, and to a method for producing desired proteins using the inventive bacteria. The invention also relates to nucleic acids coding for a SecA protein having increased levels of protein secretion and containing a gene sequence SecA or an allele, homologue or derivative of said nucleotide sequences or nucleotide sequences hybridising therewith and comprising at least one mutation. Surprisingly, just one mutation in a nucleotide of a SecA gene leads to increased levels of protein secretion or to protein secretion for the first time.

Abstract: It is an object of the present invention to provide alkaline proteases having industrially sufficient protein productivity and a significant detergency. In the alkaline proteases, the amino acid residues at (a) position 9, (b) position 49, (c) position 194, (d) position 212, (e) position 237, (f) position 245, (g) position 281, (h) position 313, (i) position 379 and (j) position 427 in SEQ ID NO: 2 are selected from the following amino acid residues; Position (a); glutamine, Position (b); glutamine, Position (c); lysine or arginine, Position (d); arginine, asparagine or glutamine, Position (e); asparagines, Position (f); asparagines. Position (g); arginine, Position (h); asparagines, Position (i); lysine, arginine, glutamic acid or aspartic acid, and Position (j); arginine.

Abstract: The present invention provides a DNA encoding a human Tumor Antigen Derived Gene-14 (TADG-14) protein selected from the group consisting of: (a) isolated DNA which encodes a TADG-14 protein; (b) isolated DNA which hybridizes to isolated DNA of (a) above and which encodes a TADG-14 protein; and (c) isolated DNA differing from the isolated DNAs of (a) and (b) above in codon sequence due to the degeneracy of the genetic code, and which encodes a TADG-14 protein. Also, provided is a vector capable of expressing the DNA of the present invention adapted for expression in a recombinant cell and regulatory elements necessary for expression of the DNA in the cell.

Abstract: The invention relates to a method of identifying a modulator of RHBDL4, said method comprising (i) providing a first and second sample of cells; (ii) contacting said first sample of cells with a candidate modulator of RHBDL4; (iii) measuring epidermal growth factor receptor (EGFR) transactivation in said first and second samples of cells, wherein a difference between the transactivation measured in said first and second samples of cells identifies said candidate modulator of RHBDL4 as a modulator of RHBDL4. The invention also relates to RHBDL4 protease assays and to uses of RHBDL4 protease and methods of cleavage of RHBDL4 substrates.

Abstract: The present invention relates to a crystal. In particular the present invention relates to a crystal of the N-domain of ACE protein. The present invention also relates to methods, processes, domain specific modulators, pharmaceutical compositions and uses of the N-domain crystal and the structure co-ordinates thereof.

Abstract: The invention provides isolated SUMO-specific protease-like (or “SSP”) domain-containing polypeptides from microorganisms, including bacteria, protozoans and yeast, including Escherichia, Salmonella, Pseudomonas, Chlamydia, Plasmodium, Trypanosma, Mesorhizobium, Rickettsia, Cryptosporidium and Candida species. The invention further provides modifications of such polypeptides, functional fragments therefrom, encoding nucleic acid molecules and specific antibodies. Also provided are methods for identifying polypeptides and compounds that associate with or modulate the activity of the SSP domain-containing polypeptides. Further provided are methods of modulating a biological activity in a cell, and treating pathological conditions, using the described nucleic acid molecules, polypeptides and compounds.

Abstract: Disclosed are the simian T-cell lymphotropic virus type 3 subtype D (STLV-3 subtype D), isolated nucleic acid molecules encoding STLV-3 subtype D polypeptides, such as STLV-3 subtype D envelope, protease, polymerase, tax, rex, and capsid polypeptides, isolated polypeptides encoded by such nucleic acids. Methods are also disclosed for detecting STLV-3 subtype D, for example by detecting a STLV-3 subtype D nucleic acid or polypeptide in the sample. Accordingly, probes, primers, and antibodies for use in detecting STLV-3 subtype D nucleic acids or polypeptides are disclosed. Therapeutic compositions which included isolated nucleic acid molecules encoding a STLV-3 subtype D polypeptides or isolated polypeptides encoded by such nucleic acid molecules are also disclosed.

Abstract: The present invention encompasses isolated human coagulation Factor VII variants comprising a substitution of Phe in position 374 of SEQ ID NO 1 with another amino acid residue.

Abstract: A protein of amino acid sequence SEQ ID NO:6 isolated from Coprinus clastophyllus. The protein is stable at 30 to 37 degrees centigrade and has an optimal prolyl oligopeptidase activity at about 45 degree centigrade and at about pH 7. The present invention also relates to a medicament comprising such protein for treating celiac disease caused by proline abundant gluten.

Abstract: The present disclosure provides crystals of and structural coordinates of Hepatocyte growth factor activator with and without bound pseudo-substrate or inhibitor. In a specific embodiment, a crystal structure of activated HGFA complexed with a Kunitz domain inhibitor is provided. The crystals and crystal structures are useful, for example, in the design and synthesis of inhibitors of HGFA.

Abstract: Stable compositions comprising a nucleic acid binding support material comprising a substrate with functional groups attached to the substrate; a lysing enzyme; a water dispersible matrix material; and a saccharide, methods of using the compositions, and products and devices which include the compositions are disclosed.

Abstract: The present invention relates to a method for producing an acidified milk drink using an enzyme which reduces the isoelectric point of the milk proteins. The invention also relates to a novel enzyme having deamidase activity and its use in production of an acidified milk drink.

Abstract: The present invention relates to structural studies of dipeptidyl peptidase I (DPPI) proteins, modified dipeptidyl peptidase I (DPPI) proteins and DPPI co-complexes. Included in the present invention is a crystal of a dipeptidyl peptidase I (DPPI) and corresponding structural information obtained by X-ray crystallography from rat and human DPPI. In addition, this invention relates to methods for using structure co-ordinates of DDPI, mutants hereof and co-complexes, to design compounds that bind to the active site or accessory binding sites of DPPI and to design improved inhibitors of DPPI or homologues of the enzyme.

Abstract: A method for high-throughput identification of antigens is disclosed. The method involves generating transcriptionally active PCR (TAP) products of one or more antigen candidates and expressing the TAP products in an in vitro translation transcription (IVTT) system. The TAP products are purified using identifiable tags. The purified TAP products are presented to isolated antigen-presenting cells (APCs), which are in turn are presented to T-cells. The ability of the antigen candidates to induce activation of the T-cells is determined. Activation of the T-cells identifies the antigen candidate as an antigen. Immunogenic compositions and methods of treatment using such compositions are also disclosed.

Abstract: A method of screening a compound which modulates dipeptidyl peptidase I (DPPI) activities comprises the steps of adding a peptide substrate of DPPI to a reaction mixture which comprises DPPI and a compound, wherein the peptide substrate of DPPI has at least 3 amino acids and binds to a binding site of DPPI in addition to the S1-S2 site; and measuring the molecular weight of the substrate, wherein a change in the molecular weight of the substrate is indicative of the presence of DPPI activity.

Abstract: Prostatic kallikrein for the manufacture of a diagnostic or pharmaceutical composition for diagnosis/treatment of type 1 allergy, especially allergy to dogs.

Abstract: The present invention is directed to the use of a compound stimulating deubiquitinating activity in a cell for the manufacture of a medicament for enhancing the expression of integral membrane proteins on the cell surface. Especially, the invention is directed to the use of such compound for the manufacture of a medicament for the treatment of a disease of condition selected from the group consisting of cystic fibrosis, diabetes insipidus, hypercholesterinaemia and long QT-syndrome-2.