62 results about "THROMBIN HUMAN" patented technology

The invention belongs to the technical field of biomedical materials, and discloses microgel assembly powder capable of rapidly stopping bleeding as well as a preparation method and application of the microgel assembly powder. The microgel assembly powder is composed of microgel powder and assembly agent powder, and the mass ratio of the microgel powder to the assembly agent powder is (1: 10)-(10: 1); the microgel powder comprises carboxymethylated and sodium trimetaphosphate cross-linked dual-modified polymer powder, and the assembling agent powder is polyphenol modified polymer powder. The microgel assembly powder can quickly absorb moisture in blood to concentrate the blood after being in contact with the blood, meanwhile, rich ortho-phenolic hydroxyl groups on the assembly agent can immediately form a hydrogen bond network among microgel, red blood cells and platelets, and the microgel assembly powder is assembled into hydrogel with excellent mechanical strength and adhesion performance. In addition, the powder can load bioactive molecule thrombin, activate multiple blood coagulation cascade reaction and promote formation of a fibrous protein network and blood clots, and through the synergistic effect of multiple hemostasis mechanisms, the powder shows rapid and effective hemostasis performance.

A tissue regeneration multi-component biomaterial composition, including: a) a chitosan component having a number average molar mass (Mn) of about 32-47 kilodalton (kDa) and a degree of deacetylation (DDA) of about 80- 90%; b) a thrombin component present in an amount effective to accelerate the solidification of the chitosan component with a blood-derived product mixture; and c) a trehalose component present in a lyoprotectant effective amount.

The invention belongs to the technical field of polypeptide preparation and biological medicine, and particularly relates to a preparation method of Jingzhaotoxin polypeptide IX (JZTX-IX) and application of Jingzhaotoxin polypeptide IX in treatment of diabetes mellitus. A chilobrachys jingzhao toxin polypeptide IX coding gene, a 6 * His tag nucleotide sequence, a thrombin restriction enzyme cutting site nucleotide sequence and an MBP hydrotropy tag nucleotide sequence are connected to a prokaryotic expression vector for prokaryotic expression, and the expressed polypeptide is purified to obtain the chilobrachys jingzhao toxin polypeptide IX protein. The target polypeptide with high purity, complete disulfide bond folding and uniform conformation is obtained, so that the high purity and biological activity of the target polypeptide are ensured. On the basis, the treatment activity of the JZTX-IX on diabetes mellitus especially type II diabetes mellitus is found and verified, and the JZTX-IX can remarkably reduce fasting blood glucose of mice with the type II diabetes mellitus, improve glucose tolerance and insulin sensitivity, relieve polydipsia and polyuria symptoms and promote insulin secretion and islet cell proliferation.

The present disclosure is directed to human protease activated receptor 4 (PAR4) binding proteins (e.g. antibodies). In particular, anti-PAR4 binding proteins which are antagonists of human PAR4, as well as methods and uses thereof.

The invention relates to the technical field of thrombin detection, in particular to a thrombin fluorescent substrate applied to whole blood as well as a preparation method and application of the thrombin fluorescent substrate. The thrombin fluorescent substrate comprises a specific recognition peptide chain D-Phe (4-MeSO2)-Pro-Gly-Arg and a fluorescent group. The thrombin fluorescent substrate disclosed by the invention has high affinity with thrombin, high catalytic efficiency and strong anti-interference capability, and can be widely applied to a thrombin detection technology.

The invention discloses a recombinant leech polypeptide with high anticoagulant activity as well as a preparation method and application thereof. A PelB-T2Y-N47K-N52Y mutant is constructed by carrying out structural optimization on a hirudin HV2 sequence, a polypeptide product with relatively high anticoagulant activity is obtained after induced expression and nickel affinity chromatography purification are carried out in a prokaryotic expression system, and the anticoagulant activity of the polypeptide product reaches 9135.8 ATU / mg and is remarkably superior to that of an initial sequence HV2. Meanwhile, the polypeptide shows a lower IC50 value in a hydroxyl radical scavenging experiment, and has the activity of inhibiting generation of AGEs in a non-enzymatic saccharification model. Molecular docking and binding free energy analysis show that the combination of the mutant and thrombin (PDB ID: 4HTC) is more stable. The leech polypeptide disclosed by the invention has the potential of being developed into a multifunctional bioactive peptide preparation, and is suitable for the related fields of anticoagulation, antioxidation and anti-saccharification.

The invention belongs to the technical field of polypeptide preparation and biological medicine, and particularly relates to a preparation method of Jingzhaotoxin polypeptide XI (JZTX-XI) and application of Jingzhaotoxin polypeptide XI (JZTX-XI) in treatment of diabetes mellitus. A Jingzhaotoxin polypeptide XI coding gene, a nucleotide sequence of a 6 * His tag, a nucleotide sequence of a thrombin restriction enzyme cutting site and a nucleotide sequence of an MBP hydrotropy tag are connected to a prokaryotic expression vector for prokaryotic expression, and a target polypeptide which is high in purity, complete in disulfide bond folding and uniform in conformation is obtained through purification. The purity and the biological activity are relatively high. Meanwhile, the treatment activity of the JZTX-XI in the aspect of diabetes is verified, and it is found that the JZTX-XI can remarkably reduce fasting blood glucose of mice with type II diabetes, remarkably up-regulate a glucose transport regulation gene Glut2, improve glucose tolerance and insulin sensitivity, relieve polydipsia and polyuria symptoms and promote insulin secretion and islet cell proliferation.

The present disclosure provides synthetic compositions useful as inhibitors of mannan-binding lectin-associated serine protease-2 (MASP-2), including compositions that selectively inhibit MASP-2 over thrombin, as well as methods for their production and use.

The present invention discloses a method for promoting heart valve endothelialization and an artificial heart valve. The method is based on modifying circular DNA on the surface of a decellularized heart valve (DHV). A DNA hydrogel targeting thrombin is obtained on the surface of the DHV by rolling circle amplification. The DNA hydrogel not only exerts an anticoagulant function, but also acts as a heme capturer, recruiting heme from the blood. The heme is further converted into biliverdin (BV) by the heme oxygenase (HO1) loaded in the DNA, ensuring the continuous production of biliverdin, promoting anti-inflammatory immune regulation and reactive oxygen scavenging, thereby creating a regenerative immune microenvironment and promoting valve endothelialization.

The invention provides a compound of formula (I): (I) or a salt thereof, wherein R1-R2, L1-L2, A. B, C, x, and y have any of the values described in the specification, as well as compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof. The compounds are useful as thrombin inhibitors.

The application discloses a thrombin solution virus inactivation device, which comprises a pump body one and a rack, the input end of the pump body one is communicated with a liquid storage device for storing thrombin solution through a liquid inlet pipe, the output end of the pump body one is provided with a liquid outlet pipe one, a protective shell is arranged at the middle position of the rack, a light transmission cylinder for placing the thrombin solution is arranged in the protective shell, a plurality of pairs of semicircular protrusions for stirring the liquid are arranged on the inner wall of the light transmission cylinder alternately, a secondary transmission gear is fixedly arranged at one end of the light transmission cylinder, the liquid outlet pipe one penetrates through the secondary transmission gear and extends into the light transmission cylinder, a plurality of ultraviolet lamps are arranged in the annular distribution between the light transmission cylinder and the protective shell, a driving motor is arranged at the top of the rack, a primary transmission gear corresponding to the secondary transmission gear is fixedly connected to the output end of the driving motor, and the primary transmission gear and the secondary transmission gear are transmissionally connected through a transmission belt, the semicircular protrusions arranged on the inner wall of the light transmission cylinder alternately can generate a turbulence effect on the thrombin solution in the light transmission cylinder when the light transmission cylinder rotates, the solution is promoted to be stirred more uniformly, the ultraviolet light can be refracted, the optical path length is increased, the ultraviolet light is more fully contacted with the solution, and thus the virus inactivation efficiency is improved.

The invention provides a method for efficiently expressing and purifying Taq DNA polymerase by using HEK293T, and belongs to the technical field of molecular biology. According to an amino acid sequence of Taq DNA polymerase, thrombin recognition and cleavage sites, six connected His amino acid tags and IFNalpha-2 signal peptides are sequentially designed at the N end of the Taq DNA polymerase, reverse translation is performed to obtain a nucleotide sequence, codons are optimized according to the environment of HEK293T cells, and then an eukaryotic expression vector is constructed. And transfecting the eukaryotic expression vector to an HEK293T cell to obtain the target protein. The purity of the obtained Taq DNA polymerase can reach 100%, the protein yield is high, about 2 mg of Taq DNA polymerase can be obtained by 20 mL of a culture medium, and the method is economical and efficient.

This invention provides 19 F-PDA-DNA-DTPA / Gd nanosensing probe, its preparation method, and its application in fluorine magnetic resonance imaging: This nanosensing probe consists of an MR signal source, a quencher, and a response linker. The MR signal source is a high-density fluorine perfluorocarbon nanoparticle, the quencher is Gd-DTPA, and the response linker is a DNA aptamer composed of a single-stranded DNA S1 and its complementary strand S2, with nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2. As a responsive nanoparticle platform, it is only visible upon activation by a specific biological trigger. Its biological application is demonstrated by targeting thrombin with the aptamer and exhibiting a strong and selective response to thrombin in live mice.

The invention discloses a method for detecting polyvinyl chloride micro-plastic through photoelectrochemistry. Specifically, the invention develops a polyvinyl chloride micro-plastic photoelectrochemical aptamer sensor, and establishes a detection method based on the sensor, the method comprises the following steps: forming a double-chain compound by a thrombin aptamer P1 and a polyvinyl chloride aptamer P2, releasing P1 after the P2 recognizes PVC, splitting P1 under the action of exonuclease I, and detecting the PVC by virtue of the split P1. At the moment, free thrombin in the system degrades the p-aminophenol modified polypeptide to generate AP, a Bi < 3 + > / Bi < 2 + > MMCT process is triggered, enhanced photocurrent is output, AP regeneration is promoted through DI-mediated EDRR, the photocurrent is further amplified, and the sensitivity of a photoelectrochemical (PEC) sensor for detecting PVC is remarkably improved. The detection method is easy to operate, high in response speed, good in selectivity and high in sensitivity, and has good accuracy in detection of actual samples (such as cosmetics as lipsticks and liquid foundation).

Therapeutic antibodies specific for osteopontin and methods of using them for treating osteopontin-associated disorders are provided. In particular, antibodies, or antigen-binding fragments thereof, that inhibit thrombin-cleavage of osteopontin or block the activity of thrombin cleavage fragments of osteopontin are provided. Additionally, antibody conjugates and pharmaceutical compositions or formulations comprising the antibodies or antibody conjugates as well as kits including the antibodies, conjugates, or formulations are also provided.

The invention provides a method for improving the appearance of a freeze-dried antithrombin III product. The method comprises the following steps: carrying out pre-freezing treatment and sublimation drying treatment on a sample containing antithrombin III; the pre-freezing treatment comprises at least two cooling steps, and the temperature of each cooling step is-60 to-20 DEG C independently; the sublimation drying treatment comprises at least three heating steps, and the temperature of each heating step is-45 DEG C to 0 DEG C independently. The antithrombin III freeze-dried product obtained by the method disclosed by the invention meets the requirements of pharmacopoeia quality standards, and has the advantages of good molding, full appearance, no shrinkage, no collapse, no compact film layer at the top, high mechanical strength, good product stability and high market value, and is beneficial to storage and transportation.

Therapeutic antibodies specific for osteopontin and methods of using them for treating osteopontin-associated disorders are provided. In particular, antibodies, or antigen-binding fragments thereof, that inhibit thrombin-cleavage of osteopontin or block the activity of thrombin cleavage fragments of osteopontin are provided. Additionally, antibody conjugates and pharmaceutical compositions or formulations comprising the antibodies or antibody conjugates as well as kits including the antibodies, conjugates, or formulations are also provided.

The application belongs to the technical field of biotechnology, and particularly relates to a purification method of type A recombinant botulinum toxin. The purification method comprises the following steps: collecting a fermentation liquor, crushing a bacterial body to obtain a crushing liquor; adding a flocculating agent to the crushing liquor to clarify, centrifuging to remove impurities to obtain a clarified liquor; adding the clarified liquor to an ion chromatography column to perform ion chromatography to obtain an ion chromatography liquor; adding thrombin to the ion chromatography liquor to perform enzymatic cutting to obtain an enzymatic cutting liquor; adding the enzymatic cutting liquor to a hydrophobic chromatography column to perform hydrophobic chromatography to obtain a hydrophobic chromatography liquor; performing ultrafiltration concentration on the hydrophobic chromatography liquor to obtain an ultrafiltration concentrated liquor; adding the ultrafiltration concentrated liquor to a molecular sieve chromatography column to perform molecular sieve chromatography to obtain the purified type A recombinant botulinum toxin. Compared with the prior art, the purification method can effectively remove impurity proteins, does not affect the integrity of light chains and heavy chains, has an amino acid sequence coverage of 100%, and does not affect the amino acid sequence coverage of 100%.

The application relates to the technical field of biological medicine, and particularly discloses a kit for determining antithrombin III activity, which comprises a first reagent, a second reagent, a third reagent and a diluent; the first reagent at least comprises a first buffer, thrombin, a serine protease inhibitor and a first stabilizer; the second reagent at least comprises a second buffer, a chromogenic substrate and a second stabilizer; the third reagent at least comprises a third buffer, heparin, isopropyl alcohol and a third stabilizer; and the diluent at least comprises physiological saline and a fourth stabilizer. In the kit, the cost is low, the anti-interference capability is strong, the sensitivity is high, the stability is good, and the kit has a wide application prospect.

The present application belongs to the technical field of biological medicine separation and purification, and particularly relates to a leech-derived hirudin separation and purification method based on thrombin affinity magnetic microspheres and application thereof. The present application uses self-prepared leaching liquor to coarsely extract hirudin from leech powder, then selects carboxyl magnetic microspheres to prepare thrombin affinity magnetic microspheres, and further separates and purifies by using the thrombin affinity magnetic microspheres, so as to finally obtain natural hirudin with an activity of 17000 ATU / g or more. The self-prepared leaching liquor can destroy the cell wall of leech, so that the hirudin in the leech is fully dissolved, thereby providing higher efficiency for the coarse extraction of hirudin in leech powder. The prepared thrombin affinity magnetic microspheres can fully utilize the sample enrichment effect thereof, and compared with agarose microspheres and polymer microspheres, can separate and purify hirudin with higher activity and purity, has a long storage period and stable quality, can provide safe and high-quality raw materials for health food, medicine or cosmetics, and is beneficial to industrialized production and application.

This invention provides a gene encoding a coral-derived anticoagulant polypeptide and its applications, belonging to the field of bioactive peptide technology. The invention provides an anticoagulant polypeptide GcKuz1, which includes the mature peptide segment shown in SEQ ID No. 1. The polypeptide GcKuz1 of this invention exerts antithrombotic and anticoagulant effects by binding to KLKB1 in the thrombin system. This polypeptide does not cause hemolysis or cytotoxicity, and has no bleeding risk, exhibiting good biocompatibility. The anticoagulant polypeptide of this invention has therapeutic effects on thrombotic diseases, such as arterial thrombotic diseases, venous thrombotic diseases, and capillary thrombotic diseases.

Novel direct thrombin inhibitors are provided herein, along with methods for their use as anticoagulants. The direct thrombin inhibitors described herein are useful in treating and / or preventing thromboembolism and bleeding or clotting disorders. Also provided herein are methods for inhibiting thrombin in a cell using the compounds and compositions described herein.

The application discloses a preparation device of autologous thrombin, and belongs to the technical field of biomedical engineering, which comprises an activator cylinder and a conducting pipe, a piston head of a piston of the activator cylinder is arranged in an inner container, the inner container is sleeved with an outer cylinder, and a piston seat of the piston is fixedly connected with the outer cylinder. When the outer cylinder is quantitatively moved upward relative to the inner container, the activator in the inner container is pushed by the piston to be added into a test tube through the conducting pipe, so that the activator can be quantitatively added into the test tube, and the added activator can be guaranteed not to be polluted by the outside. The preparation device of autologous thrombin is used to add the activator into centrifuged whole blood to make the whole blood be mixed uniformly, and after standing and contacting, a gel is formed in the mixture, supernatant is formed in the upper part, and the supernatant is sucked to prepare a thrombin solution. The application has the advantages of convenient operation, simple process, effective prevention of external infection, high activity of prepared autologous thrombin and fast gel formation.

The present application belongs to the technical field of polypeptide preparation and biopharmacy, and particularly relates to eggshell membrane polypeptides with thrombin inhibitory activity and application thereof. Two polypeptides are obtained by separation and purification from eggshell membranes, which are nonapeptide YDGQHGRVS and octapeptide EVPPSPAL. It is proved by experiments that the nonapeptide YDGQHGRVS and the octapeptide EVPPSPAL have thrombin inhibitory activity equivalent to that of the positive control heparin sodium, and it is found that the nonapeptide YDGQHGRVS prolongs activated partial thromboplastin time and prothrombin time. The nonapeptide YDGQHGRVS and the octapeptide EVPPSPAL provided by the present application are derived from eggshell membranes, are natural in origin, have good thrombin inhibitory activity, high safety, and have good application prospect.

The invention belongs to biomedical materials, and particularly relates to an antibacterial and anticoagulant coating material for a blood contact material and application of the antibacterial and anticoagulant coating material. The antibacterial and anticoagulant coating material comprises copper tetracarboxyl phenyl porphyrin loaded with an anticoagulant and water-soluble polysaccharide. A coating formed on the basis of the antibacterial and anticoagulant coating material can achieve antibacterial and anticoagulant effects at the same time, for example, the bacteriostasis rate on escherichia coli and the bacteriostasis rate on staphylococcus epidermidis can reach 90%; in addition, the activated part thromboplastin time, prothrombin time and thrombin time of the coating are obviously higher than those of an untreated blood contact material; and the hemolysis rate is lower than 5%.

This invention discloses an earthworm protein peptide with in vitro thrombin-inhibiting activity and its preparation method, belonging to the field of functional food technology. The method first pre-treats earthworm raw materials to obtain earthworm protein; then, it uses pepsin and alkaline protease for stepwise enzymatic hydrolysis under varying temperature and pH, and ultrafiltration to collect short peptide solutions with a molecular weight below 3 kDa; the short peptide solution is specifically captured using an affinity chromatography material coupled with bovine thrombin, followed by elution, nanofiltration desalting concentration, and spray drying to obtain the final product. This invention fully releases the peptides through a dual enzymatic hydrolysis process and utilizes thrombin affinity chromatography to directionally enrich peptides with specific inhibitory activity, effectively solving the problems of low content and poor targeting of active ingredients in earthworm protein peptides in existing technologies, and significantly improving the purity and in vitro anticoagulant activity of the product.

The invention discloses an application of Spy Tag / Spy Catcher in preparation of a thrombin-antithrombin III compound, a preparation method of the thrombin-antithrombin III compound and an application of the thrombin-antithrombin III compound. The preparation method comprises the following steps: respectively obtaining a Spy Catcher-Thrombin gene segment and a Spy Tag-Antithrombin gene segment; respectively connecting the gene segments to starting vectors to form corresponding connectors; carrying out conversion and screening operation on the linker to obtain a corresponding recombinant plasmid; carrying out transformation and induction operation on the recombinant plasmid, so as to obtain a Spy Catcher-Thrombin recombinant protein and a Spy Tag-Antithrombin recombinant protein; the preparation method comprises the following steps: mixing the Spy Catcher-Thrombin recombinant protein with the Spy Tag-Antithrombin recombinant protein, and carrying out a reaction, so as to obtain the TAT protein. According to the method, the spontaneous reaction of Spy Tag / Spy Catcher is utilized, other chemical cross-linking agents are not needed, the cost can be reduced, green production is facilitated, and the recombinant protein can be produced on a large scale by utilizing a cell culture technology; the recombinant TAT antigen and the recombinant TAT antibody prepared by the invention have strong immunoreactivity, and completely meet the requirements of thrombus detection on the TAT antigen.

This invention proposes a tPA delivery system based on a tetrahedral DNA origami structure, its preparation method, and its applications, relating to the biomedical field. It includes a carrier composed of a tetrahedral DNA origami structure, a thrombus-targeting peptide, and short-chain DNA. The thrombus-targeting peptide and short-chain DNA are pre-coupled to form a peptide-short-chain complex, which is grafted onto the tetrahedral DNA origami structure. tPA and short-chain DNA are pre-coupled and loaded onto the edges of the tetrahedral DNA origami structure. Using the tetrahedral DNA origami structure as the core carrier, the vertices can be precisely modified with thrombus-targeting peptides to achieve targeting of the thrombus site. The tPA molecule is loaded onto the edges of the DNA origami structure via sulfo-SMCC linkage, and its release is controlled by introducing a thrombin-responsive DNA locking mechanism. This invention overcomes the limitations of current tPA thrombolytic therapy, enabling on-demand release of tPA during treatment, achieving local thrombolysis while reducing the risk of systemic bleeding.

This invention relates to the field of balloon technology, and discloses a functional balloon, its preparation method, and its application. The surface of the functional balloon is loaded with polyglutamic acid, which is linked to a DNA hydrogel via amide bonds. A nucleotide sequence is linked to the DNA hydrogel and thrombin. The balloon obtained by this invention can achieve hemostasis and reduce bleeding symptoms without vascular occlusion.