26 results about "THROMBIN HUMAN" patented technology

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 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 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.

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.

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.

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 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.

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.

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.

The invention discloses a vector for expressing snake venom thrombin-like bacteria, yeast engineering bacteria and a preparation method, and belongs to the technical field of gene engineering. Based on pPIC9K, pAO815 alpha, thrombin-like snake venom agkihpin and a pichia pastoris expression system, the co-expression copy number of the agkihpin gene and an auxiliary factor is optimized, the copy number of an auxiliary gene Bmh2 is adjusted, and culture parameters such as the initial pH of a culture medium, the methanol addition amount, the induction time, the induction temperature and an auxiliary agent are improved by the system, so that the expression level of the agkihpin is effectively improved, and the expression level of the agkihpin is improved. And a solid foundation is laid for agkihpin industrial application of the thrombin-like snake venom.

The utility model discloses a novel thrombin time determination kit, which relates to the technical field of chemical analysis and comprises a kit body and a kit cover, the kit cover is arranged at the top end of the kit body, first fixing frames are symmetrically mounted on the front side and the back side of the kit body, and second fixing frames are symmetrically mounted on two sides of the kit body. Fixing assemblies are symmetrically arranged on each first fixing frame and each second fixing frame, each fixing assembly comprises fixing cylinders symmetrically mounted on the corresponding first fixing frame and the corresponding second fixing frame, and by arranging the first fixing frames, the second fixing frames, the second placing frames and the fixing assemblies, a light-proof plate can be placed on the outer side of the kit body; according to the kit, the light-proof plate is fastened and limited, the light-proof plate can shield the kit body on the outer side when the kit body is stored or transported, damage to reagents in the kit caused by sunlight irradiation is avoided, and the kit cover can be conveniently closed and opened at the top end of the kit body through the arranged first spring and the arranged clamping block.

A thrombin-responsive coacervate composition comprising a coacervate comprising YR-based peptides having a thrombin cleavage sequence, an anticoagulant, and a polyphenol to stabilize the coacervate structure; and methods of using the thrombin-responsive coacervate to selectively release an anticoagulant only when thrombin is present.

This invention discloses a method for preparing a D-dimer and FDP composite quality control sample, belonging to the field of diagnostic reagent technology. The method includes the following steps: Step 1: Weigh the cryoprecipitate, add it to buffer solution, melt it in a 37°C water bath, and collect the supernatant by centrifugation; Step 2: Add trypsin, calcium chloride solution, and thrombin, mix well, and let it stand in a 37°C water bath; Step 3: Detect the FDP and DD content using an automated coagulation analyzer; Step 4: Terminate the reaction with a stop solution; Step 5: Prepare the composite quality control sample according to the target content and ratio of DD and FDP. This method is simple, low-cost, and yields extremely high concentrations of D-dimer and FDP. The prepared D-dimer and FDP composite quality control sample has an adjustable ratio, good stability, and broad applicability to reagents and instruments.

The invention belongs to the field of basic medicine, and particularly relates to a method for establishing a blood coagulation disorder animal model, which comprises the following steps of: performing intragastric administration on a healthy rat with a preparation containing podophyllotoxin according to the dose of 10mg / kg per day, and continuously performing administration for 4 days; wherein the drug administration can induce the rat to generate blood coagulation disorder phenotypes, and the phenotypes at least comprise prothrombin time prolonging, activated partial thromboplastin time prolonging and thrombin time prolonging. According to the establishment method of the blood coagulation disorder animal model, it is determined that podophyllotoxin can cause whole-body blood coagulation disorder, and the stable and repeatable blood coagulation disorder animal model is provided.

The present disclosure relates to a method for preparing a haemostatic composition comprising thrombin, the method comprising the step of reconstituting a dry thrombin directly in a paste, such as a paste comprising a biocompatible polymer. The haemostatic composition comprising thrombin may be prepared from a dry thrombin composition and a paste in a single step operation and be used for treatment of a wound.

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 discloses an extraction and separation method of anise polysaccharide with anticoagulant activity. The method comprises the following steps: crushing fresh anise, extracting with hot water, concentrating a water extract, and adding ethanol for precipitation to obtain crude polysaccharide; removing protein by adopting a Sevage method; and dialyzing to remove small molecular impurities, and freeze-drying to obtain the purified anise polysaccharide. The molecular weight of the polysaccharide is about 72.053 kDa. In-vitro coagulation experiments show that the polysaccharide can significantly prolong prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT) and reduce the content of fibroblast egg (FIB) white, and shows significant anticoagulant activity. The star anise polysaccharide provided by the invention can be used for preparing anticoagulant drugs, and has the advantages of wide raw material source, simple extraction process, definite activity and the like.

This invention relates to a recombinant hirudin fusion protein with collagen affinity, its preparation method, and its applications. The fusion protein is a recombinant hirudin fusion protein incorporating a collagen-binding domain (CBD). The fusion protein of this invention possesses affinity for type I and type III collagen, and can inhibit thrombin activity to exert an anticoagulant effect. Furthermore, it can bind well to collagen-based composite materials, making it suitable for use in cardiovascular implants such as heart valves, cardiac occluders, dialysis membranes, oxygenation membranes, artificial blood vessels, and vascular stents, as well as in extracorporeal circulation and blood purification devices, enhancing anticoagulant and antithrombotic effects.

The present application relates to a kind of whole-cell biosensors of E. coli surface display, for realizing the high sensitivity detection of multiple proteases.The sensor is displayed on the surface of E. coli by synthetic biology method and includes anchor module, protease-specific cleavage peptide and fluorescent reporter module of recombinant fusion protein.When target protease exists, its specific cleavage peptide causes fluorescent reporter module to dissociate from cell surface, and fluorescence signal is weakened, and the degree of signal attenuation is positively correlated with protease concentration and activity.The detection sensitivity of the present application is high, and the detection limit of matrix metalloproteinase-2, thrombin and other proteases reaches the level of femtogram per milliliter, which is more than three orders of magnitude higher than traditional methods;It is simple to operate, only needs to be added sample-incubation-detection three steps, and the cost of single detection is less than 0.1 dollar;It can directly detect protease activity in complex biological matrix without expensive antibody, and has good clinical application and large-scale screening potential.