69 results about "Zymogen" patented technology

The application belongs to the technical field of genetic engineering, and discloses a kind of saccharomyces cerevisiae for producing rare ginsenoside by seaweed biomass and a construction method and application thereof.The saccharomyces cerevisiae has the following characteristics: overexpression of agarase, neojuncanohydrolase, hydroxymethylglutaryl coenzyme A reductase, isopentenyl diphosphate delta isomerase, dammaradienol synthase, protopanaxadiol synthase, cytochrome P450 reductase, protopanaxatriol synthase and glycosyltransferase.The application combines enzymolysis of seaweed biomass with fermentation of rare ginsenoside, which not only endows wild-type yeast with the ability to degrade seaweed biomass that it originally does not have, but also effectively improves the yield of squalene and downstream terpenes (rare ginsenoside Rh1) by overexpression of tHMG1 and IDI1.The saccharomyces cerevisiae can effectively utilize seaweed biomass to obtain squalene and rare ginsenoside Rh1, and has the characteristics of simplicity, economy and ecological friendliness, can convert cheap biomass into high-value products, and provides a way for the development of marine bioeconomy.

The present disclosure relates to enzymes, compositions and methods for performing conditional homologous recombination of a targeted DNA molecule or genome by using modified proteins comprising nucleic acid DNA binding proteins with protease-conditional recombinase (hereinafter “ProRec”) activity. Namely a new cre recombinase pro enzyme, compositions / kit and sensors comprising the cre recombinase pro enzyme of the present disclosure. Namely, a cre recombinase pro-enzyme, for identifying and quantifying proteolytic activity in a disease or an infection, comprising intein mediated circularization, a flip-excision cassette comprising an output reporter and / or effector protein; a linker sequence cleavable by a protease.

This invention discloses a fusion protein targeting the urokinase-type plasminogen activator receptor and its application in the field of anti-aging. The fusion protein is composed of a hepatocyte growth factor active domain and a humanized anti-uPAR single-domain antibody linked by a flexible linker peptide, and its specific amino acid sequence is SEQ ID NO:3. This fusion protein can specifically bind to uPAR, which is highly expressed on the surface of senescent cells, with intramolar affinity, achieving targeted delivery of the hepatocyte growth factor active domain. In vitro experiments have demonstrated that this protein can significantly reverse the aging phenotype of human fibroblasts, effectively reduce aging-related β-galactosidase activity, p16INK4a protein expression, and interleukin-6 secretion, and promote cell proliferation. In a rapidly aging mouse model, this protein can systematically improve age-related physiological functional decline. The fusion protein of this invention achieves specific targeting and efficient treatment of senescent cells, providing a new solution for the development of anti-aging drugs.

To provide methods of identifying new substrates for proteases and methods of using these substrates in a variety of therapeutic, diagnostic and prophylactic indications.SOLUTION: The invention relates generally to: polypeptides that include a cleavable moiety that is a substrate for at least one protease selected from matriptase and u-plasminogen activator (uPA); activatable antibodies and other large molecules that include the cleavable moiety that is a substrate for at least one protease selected from matriptase and u-plasminogen activator; and methods of making these polypeptides that include the cleavable moiety that is a substrate for at least one protease selected from matriptase and u-plasminogen activator, and methods of use in a variety of therapeutic, diagnostic and prophylactic indications.SELECTED DRAWING: Figure 4A

The present application relates to a method for promoting the degradation of pathological TDP-43 protein, comprising administering to a subject a therapeutically effective amount of one or more compounds selected from components of the plasminogen activation pathway, compounds that can activate plasminogen directly or indirectly by activating upstream components of the plasminogen activation pathway, compounds that mimic the activity of plasminogen or plasmin, compounds that can upregulate the expression of plasminogen or plasminogen activators, plasminogen analogs, plasmin analogs, tPA or uPA analogs, and antagonists of fibrinolysis inhibitors. The present application also relates to agents that promote the degradation of pathological TDP-43 protein and uses thereof.

Disclosed herein, are peptides that bind urokinase-type plasminogen activator receptors. The peptides may comprise amino acid sequences of IPPWEAPK (SEQ ID NO: 1), DLAQCQTPTQAAPPTPVSPR (SEQ ID NO: 2), or LHVPLMPAQPAPPK (SEQ ID NO: 3), or retro-inverso amino acid sequences of the above enumerated sequences. Also described herein, are methods of administering compounds comprising peptides that bind urokinase-type plasminogen activator receptors to subjects for the treatment of ovarian cancer and improving wound closure.

The current invention relates to a polypeptide comprising an immunoglobulin single variable domain and an urokinase-type plasminogen activator (uPA) domain wherein the immunoglobulin single variable domain is able to bind to fibrin via a fibrin-binding domain.

The invention discloses a polypeptide asparagine ligase VdiPAL1 and an application of the polypeptide asparagine ligase VdiPAL1. The polypeptide asparagine ligase VdiPAL1 is newly found from viola plants, 12 mg / L zymogen protein is obtained through expression of an escherichia coli prokaryotic system, and compared with homologous enzymes VyPAL2 and VdiPAL1 which are widely applied at present, the polypeptide asparagine ligase VdiPAL1 has the advantages of high expression, high catalytic efficiency and substrate specificity and has good development and application potential. The invention also shows the value of the VdiPAL1 for guiding homologous enzyme modification, and particularly discloses the mutant protein VyOPT of the polypeptide asparagine ligase VyPAL2 and a coding cDNA (complementary deoxyribonucleic acid) sequence of the mutant protein VyOPT. The invention also discloses a mutant VyOPT, which comprises five mutations obtained by modification guided by the VdiPAL1. Compared with the expression quantity of the natural VyPAL2, which is about 0.1 mg / L, the expression quantity of the VyOPT reaches 12 mg / L.

The present invention provides methods for modifying a donor organ, e.g., liver, or a portion thereof. The methods comprise contacting the donor liver with lipid nanoparticles comprising RNA. Compositions of lipid nanoparticles comprising RNA are also provided. The disclosed LNPs may comprise an siRNA, e.g., targeting tissue plasminogen activator (tPA) or plasminogen activator inhibitor 1 (PAI-1). The LNPs may comprise an mRNA encoding a protein, e.g., Nuclear Factor-erythroid factor 2-related factor 2 (NRF2) or thrombomodulin.

The invention relates to the technical field of membrane separation, and discloses a high-protein milk concentration process based on a membrane filtration technology, the process adopts an electric response membrane grafted with an electric response polymer brush on the surface, and an electric control zymogen modified by genetic engineering is added into milk feed liquid to be treated. The zymogen is modified with an electric field responsive anchoring group and an electric signal activated ferrocene protecting group. In the filtering stage, a first electric field is applied to stretch the polymer brush, so that anti-pollution filtering is realized; and in the cleaning stage, switching to a second electric field to enable the polymer brush to shrink and drive the zymogen to be anchored on the surface of the membrane, and applying an activation pulse to trigger the activity of the zymogen so as to degrade pollutants in situ. On the premise of not interrupting production, not adding chemical cleaning agents and not damaging target product protein, milk concentration is efficiently and continuously carried out, and long-term stable high permeation flux is maintained.

The present invention relates to double stranded ribonucleic acid (dsRNA) compositions targeting the plasminogen (PLG) gene, as well as methods of inhibiting PLG expression, and methods of using such dsRNA compositions to treat subjects that would benefit from reduced PLG expression, such as subjects having a PLG-related disease, disorder or condition.

The invention discloses a PAI-1 protein targeting peptide compound (called P peptide for short) with a thrombus dissolving effect and application thereof, and belongs to the field of biological medicine. The PAI-1 targeted thrombolytic polypeptide compound comprises P1, P2, P3 and P4, and is characterized in that the compound is derived from a core structure 'X-Asp-Glu-Leu-Cys-X-Tyr-Tyr-X-X-Cys-X' (cyclization is performed through a disulfide bond between Cys, and X represents any type of amino acid), and the compound can be specifically bound with a key binding site of PAI-1. The polypeptide can significantly reduce the inhibitory effect of PAI-1 on plasminogen activators (tPA, uPA), enhance the activity of a fibrinolytic system, and generate in-vivo and in-vitro thrombolysis effects. The polypeptide compounds P1, P2, P3 and P4 not only have a thrombolysis effect, but also can reduce the expression level of pro-inflammatory cytokines (IL-1beta, TNF-alpha, IL-6 and the like), which shows that the polypeptide compounds also have an anti-inflammatory effect. In conclusion, the polypeptide compound can be used for preparing a PAI-1 antagonist, a thrombolytic agent and an anti-inflammatory agent related to thrombus.

This invention relates to the field of biological detection, and more particularly to the preparation method and application of the t-PAI·C detection kit. The invention provides a detection reagent comprising: antibody-coated magnetic microparticles and enzyme-labeled antibodies; the antibodies include: tissue plasminogen activator-plasminogen activator inhibitor-1 complex antibody. In the detection reagent provided by this invention, when the sample contains the tissue plasminogen activator-plasminogen activator inhibitor-1 complex, the antibody coated with the magnetic microparticles and the enzyme-labeled antibody undergo an immune reaction with the complex, forming a sandwich-like immune complex consisting of the antibody coated with the magnetic microparticles, the complex, and the enzyme-labeled antibody; the enzyme on this complex acts on a luminescent substrate and emits light, and the concentration of the complex in the sample is calculated by detecting the luminescence signal.

A method comprising the steps of: (a) assessing an expression level of at least one protein, selected from Table 1, 2 or 3 in a sample from a subject, where in the at least one protein may be selected from the list comprising: Beta-Ala-His dipeptidase, Apolipoprotein L1, Methanethiol oxidase, Vitamin K-dependent protein S, von Willebrand factor, Plasminogen, Selenoprotein P, Protein disulfide-isomerase A6 and Inter-alpha-trypsin inhibitor heavy chain H3, and (b) using the expression level to determine whether the subject has endometriosis.

The application discloses an engineered exosome targeting human urokinase-type plasminogen activator receptor uPAR and a preparation method and application thereof. Firstly, a high-affinity anti-human uPAR single-domain antibody VHH is prepared; then, a human umbilical cord mesenchymal stem cell engineering cell line expressing a VHH-PTGFRN fusion protein is constructed, and the engineered exosome is prepared and purified by combining three-dimensional hypoxic culture, chemical induction and size-exclusion chromatography technology. The engineered exosome comprises four comparative samples, wherein the core sample Exo^(VHH / miR) can simultaneously display the anti-uPAR VHH on the membrane surface to realize targeted delivery and enrich miR-193b-3p in the chamber to realize gene silencing, and the synergistic effect of the two can efficiently remove the uPAR high-expression senescent cells and inhibit the senescence-associated secretory phenotype SASP. The engineered exosome has the advantages of standardized preparation process, high yield and high purity, strong targeting, excellent curative effect and good safety, and can be used for preparing a treatment preparation for anti-aging and aging-related diseases, and has important clinical application value.

To provide compositions and methods for modulating the plasmin system for treating and / or preventing various pathological conditions.SOLUTION: The present invention relates to antigen binding proteins comprising an antigen binding domain that binds to plasminogen, wherein the antigen binding protein reduces the activation of plasminogen, pharmaceutical compositions comprising the same, and methods and uses thereof.SELECTED DRAWING: None

This invention belongs to the field of biomedical technology, specifically relating to a recombinant human plasminogen mutant, its preparation method, and its application. A recombinant human plasminogen mutant, the amino acid sequence of which is shown in SEQ ID NO.3. The recombinant human plasminogen mutant of this invention, compared to wild-type mPLG, can be directly expressed solublely in the supernatant of *E. coli* fermentation broth. Soluble expression allows for direct chromatography of the fermentation broth to obtain the target protein, which has significant advantages over inclusion body renaturation treatment in terms of process simplification, activity loss, time cost, and production cost. Furthermore, the activity of the mutant protein was tested using the fibrin plate method and kit method. The results showed that the titer of the target protein was approximately 8 UI / mg, which is similar to the titer of mPLG obtained by renaturation before mutation and PLG obtained from human blood. In summary, compared to the wild type, the mutant strain exhibits significantly improved solubility while maintaining high protein activity.

The invention discloses a kit for detecting susceptibility and severity of children acute respiratory distress syndrome, and relates to the technical field of biomedical detection. The kit comprises a detection reagent combination and a standardized reference system, the detection reagent combination is prepared from specific binding angiogenin 2, a soluble advanced glycosylation end product receptor, pulmonary surfactant protein D, a soluble tumor necrosis factor receptor 1, interleukin 6, interleukin 8, a plasminogen activator inhibitor 1 and a capture and detection antibody of III type procollagen N-terminal peptide. The primer pair and the probe are used for specifically amplifying a single nucleotide polymorphism site rs4073 in a promoter region of the interleukin 8 gene; the standardized reference product system comprises gradient standard products covering the physiological concentration dynamic ranges of the markers on the 0th day, the 3rd day and the 7th day of morbidity; the kit is used for early-stage precise layering and dynamic prognosis evaluation of the acute respiratory distress syndrome of children.

To provide an improved method for treating IdiopathicPulmonaryFibrosis (IPF).SOLUTION: The invention features a polypeptide comprising a variant of plasminogen-activator inhibitor-1 (PAI-1) having a reduced capacity to bind vitronectin, a reduced capacity to interact with the PAI-1 clearance receptor LDL-receptor-related-protein 1 (LRP1), and a capacity to efficiently inhibit neutrophil elastases in presence of neutrophil extracellular traps (NETs). In some embodiments, a polypeptide of the present invention comprises a PAI-1 variant optionally fused to an Fc domain monomer or moiety. The invention also features pharmaceutical compositions and methods of using the polypeptides to treat diseases and conditions characterized by aberrant neutrophil elastase activity (as an example, idiopathic pulmonary fibrosis).SELECTED DRAWING: Figure 1

This disclosure relates to nanoparticles coated with fusion proteins comprising a domain that binds a cancer marker and a domain comprising a toxic polypeptide. In certain embodiments, the targeted cancer marker is urokinase plasminogen activator receptor (uPAR) insulin-like growth factor 1 receptor (IGF1R), EGFR, HER2, and / or other member of the ErbB family of receptors. In certain embodiments, the molecule that binds a cancer marker is an amino terminal fragment of uPA or variant capable of binding uPAR and / or IGF1 or variant capable of binding IGF1R. In certain embodiments, the toxic polypeptide is a bacterial exotoxin.

The invention discloses a preparation method and application of recombinant pepsin, and belongs to the technical field of enzyme engineering. The invention mainly aims to provide a method for commercially preparing recombinant pepsin and application of the recombinant pepsin, so that animal-derived pepsin is replaced by the recombinant pepsin to solve the current application problem. Specifically, the method comprises the following steps: introducing a pepsinogen coding gene containing a signal peptide as shown in SEQ ID NO.2 into pichia pastoris host bacteria to obtain recombinant pichia pastoris; performing high-density fermentation on the recombinant pichia pastoris to obtain fermentation liquor containing recombinant pepsinogen; performing ultrafiltration concentration and filter washing dialysis on the fermentation liquor to obtain a zymogen solution, and activating the zymogen solution under an acidic condition to obtain an active recombinant pepsin solution; and purifying the recombinant pepsin solution to obtain the recombinant pepsin. The zymogen produced by the method disclosed by the invention is high in yield, and the pepsin obtained from the zymogen has the same function as the extracted pepsin.

The present disclosure provides a duplex or single-stranded siRNA molecule against plasminogen, the siRNA molecule containing modified or unmodified nucleotides and wherein at least one strand of the duplex or the single-stranded siRNA has a sequence that has at least 80% sequence identity to any one of SEQ NOs: 1 to 28. Further provided is a duplex or single-stranded siRNA molecule against plasminogen, the siRNA molecule containing modified or unmodified nucleotides and is between 25 and 35 nucleotides in length. The siRNA molecule may be formulated in a lipid nanoparticle as described herein.

Disclosed are novel urokinase plasminogen activator receptor (uPAR) binding molecules and soluble urokinase plasminogen activator receptor (suPAR) binding molecules and methods of their use.

Provided is a pharmaceutical composition with which it is possible to prevent the development of late-onset cerebral ischemia and obtain a prognosis-improving effect by performing active hematoma removal using a tissue plasminogen activator. This pharmaceutical composition contains a tissue plasminogen activator as an active ingredient and is for use in hematoma removal therapy in the cerebral cistern to remove a dislodged hematoma by suction by using the tissue plasminogen activator in an aqueous solution state as a cleansing agent, said therapy being performed at the same time as treatment of a cerebral aneurysm by means of craniotomy with clipping during the acute phase of a subarachnoid hemorrhage associated with a ruptured cerebral aneurysm.

This disclosure relates to isolated oligonucleotides comprising duplex regions targeting human PLG mRNA, and delivery systems, kits and compositions comprising the same, and methods of using the same for inhibiting or downregulating PLG gene expression and treating or preventing a disease or disorder associated with aberrant or increased expression or activity of PLG or a disease or disorder where PLG plays a role.

The invention discloses a tachypleus amebocyte lysate based on a peptide-rhodamine 110 conjugate and application of the tachypleus amebocyte lysate in endotoxin detection. The tachypleus amebocyte lysate comprises a C factor, a B factor, a clotting zymogen and a peptide-R110 conjugate. The peptide-R110 conjugate is adopted as a limulus reagent substrate, when the limulus reagent substrate is applied to endotoxin detection, a fluorescence detection background signal can be remarkably reduced, the detection sensitivity is greatly improved, meanwhile, the dual detection functions of color development and fluorescence are achieved, end-point method reading can be achieved in a fluorescence mode, and the detection sensitivity is greatly improved. High-precision and high-reliability detection of trace endotoxin is realized, and the method has important significance in quality control in the pharmaceutical production process, safety evaluation of medical equipment and other fields needing endotoxin detection.

The present disclosure provides combinations of tissue plasminogen activator mutant / s and lysine analogs and compositions thereof for inhibiting fibrinolysis induced by free plasmin. The present disclosure further provides kits, uses and methods for treating coagulopathy associated with anti-fibrinolytic therapy.