16 results about "Thrombin activity" patented technology

Disclosed are a fibrinogen solution and a thrombin solution. The fibrinogen solution comprises fibrinogen at a concentration of at least 40 mg / ml, factor XIII, pharmaceutically acceptable additives and water. The dynamic viscosity of the fibrinogen solution measured at 20° C. increases at most by 35% after storing the solution at 20° C. for 30 days. The thrombin solution comprises thrombin, pharmaceutically acceptable additives and water. The thrombin activity decreases at most by 15% after storing the solution at 25° C. for 14 days. Also disclosed is a fibrin sealant kit with a first container comprising the fibrinogen solution and a second container comprising the thrombin solution. Further, methods for preparing a fibrin sealant and methods for treating a wound are disclosed.

Provided herein are method and kits for identifying thrombus origin based on temporal thrombin activity assay, and use thereof for optimizing post-ischemic event treatment, and further preventing recurrence of secondary ischemic events.

The invention belongs to the technical field of nano preparations, and particularly relates to a hirudin nano drug for relieving chronic asthma and a preparation method thereof.The hirudin nano drug is prepared from, by weight, 0.5-1 part of hirudin prodrug, 10-15 parts of stabilizer, 40-80 parts of freeze-drying protective additive and 1.5 parts of histidine; the hirudin prodrug molecule is composed of hirudin, MMP-9 responsive peptide, ROS responsive thioether bond and hydrophobic hexadecanoic acid, can spontaneously assemble to form a nano-sized prodrug under hydrophilic and hydrophobic effects, has stability in systemic circulation and is targeted and enriched at an asthma airway inflammation part; the hirudin with the thrombin activity inhibition effect is responsively released at the focus, so that the effect and the purpose of slowly treating the asthma are achieved.

The invention belongs to the technical field of anticoagulant drugs, and particularly relates to lumbricus protein with anticoagulant activity as well as a preparation method and application of the lumbricus protein. The lumbricus protein provided by the invention contains at least one of a c71616 protein with an amino acid sequence as shown in SEQ ID No. 1, a c128686 protein with an amino acid sequence as shown in SEQ ID No. 2, a c58213 protein with an amino acid sequence as shown in SEQ ID No. 3 and a c118923 protein with an amino acid sequence as shown in SEQ ID No. 4. The protein can effectively inhibit thrombin activity and interfere formation of fibrin so as to act on a key link of a blood coagulation process. The protein can be obtained through recombinant expression and purification, and has high expression efficiency and stability, so that the protein can be used as an active component to be applied to development of anticoagulant drugs, surface anticoagulant coatings and other products.

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.

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 a method for detecting thrombin activity in a biological product by using a functionalized electrode, and belongs to the technical field of electrochemical detection. Firstly, thrombin specific substrate polypeptide is fixed on the surface of a silk-screen printing gold electrode through an Au-S bond, and the N end of the polypeptide is modified with ferrocene (Fc) to serve as an electroactive signal molecule; then the surface of the electrode is coated with a cellulose acetate membrane (CA) to obtain a functionalized electrode, and the CA membrane can reduce non-specific adsorption and prevent electrode pollution and signal attenuation; dropwise adding a solution containing thrombin onto the functionalized electrode for reaction; and washing the reacted electrode, testing by using a square wave voltammetry, and obtaining the thrombin activity according to the change of the peak current. The method is convenient to operate, high in specificity, short in consumed time and long in storage time of the functionalized electrode, and is an effective means for detecting the thrombin activity in the biological product.

The invention relates to a method for rapidly detecting the activity of human thrombin in a human prothrombin complex, and belongs to the technical field of biological product detection. The method comprises the following steps: diluting a human thrombin solution into a plurality of gradients in equal proportion, then adding a fibrinogen solution, standing for a period of time, observing whether clots or fibrin are separated out, and determining the corresponding human thrombin activity titer limit when the clots or fibrin are separated out; mixing the human prothrombin complex with a fibrinogen solution, and detecting the refractive index under the wavelength of 589 nm; and determining the activity of the human thrombin in the human prothrombin complex by comparing the refractive index values. According to the method, the original 24-hour test time is shortened to be within 1 hour, and the problem that the time of an existing method for testing the activity of the human thrombin in the human prothrombin complex is too long is solved.

The invention discloses a hirudin separation and purification method based on affinity chromatography, and belongs to the technical field of hirudin separation and purification. Aiming at the characteristics of high viscosity and many impure proteins of a hirudin crude extract, an affinity chromatography medium polystyrene microsphere is modified: modified methoxy polyethylene glycol-polyaspartic acid containing 3, 5-di-tert-butyl-4-hydroxycinnamic acid is grafted, so that the hydrophilicity is improved, the non-specific adsorption is reduced, and the affinity chromatography efficiency is improved; meanwhile, free radicals are removed by utilizing a hindered phenol structure so as to protect the activity of thrombin; after hydroformylation treatment, fixing thrombin as a ligand to form a stable affinity medium; and combining with thrombin affinity chromatography separation, and then carrying out ultrafiltration concentration, desalination and freeze drying to obtain hirudin. A macroporous structure is reserved through a modified medium, the mass transfer resistance is reduced, and the binding efficiency of hirudin and ligands is improved; the activity loss of the hirudin is reduced in a subsequent process. The method can efficiently separate and purify hirudin, has good adsorption selectivity and stability, and is suitable for large-scale separation.

The invention provides a powerful hemostatic adhesive prepared by taking American cockroach dregs as a main raw material. The preparation method comprises the following steps: firstly, extracting and preparing chitin whiskers from waste American cockroach dregs, taking the chitin whiskers as a polymer matrix of the adhesive, taking an aqueous solution of a deep eutectic solvent as a system medium, taking thrombin as a hemostatic functional substance, and improving the overall mechanical properties by gelatin, thereby finally obtaining the biological adhesive with good adhesion and hemostatic performance. According to the product, under the condition that a cross-linking agent does not need to be used, through heating-cooling operation, a polymer skeleton network is constructed through interaction between chitin whiskers and gelatin, a channel is provided for transmitting moisture and nutrient substances to damaged tissues, and the product has degradability while achieving an ideal adhesion effect; the whole system effectively fuses the components, is friendly and easy to prepare, and integrates respective advantages. In addition, the activity and the stability of the thrombin in the eutectic solvent are both in ideal states, so that the adhesive can play a powerful hemostatic role.

The invention relates to black date polypeptide and application thereof in synergic antithrombosis with polyphenol, belongs to the technical field of biology, and particularly relates to thrombin inhibitory peptide as well as a preparation method and application thereof. The method comprises the following steps: extracting protein by taking black date wine residues as a raw material, preparing active polypeptide by a biological enzymolysis method, analyzing and screening thrombin inhibitory peptide PPHG by utilizing bioinformatics, and achieving an IC50 value of 30.69 + / -0.54 mg / mL to thrombin. The inhibition effect of the combination of the thrombin inhibitory peptide PPHG and the gallic acid GA on the thrombin activity is further explored, when the concentration ratio of the PPHG to the GA is 5: 2, the synergistic inhibition rate of the PPHG / GA on the thrombin is 38.76% + / -2.05%, and the synergistic score of the ZIP is the highest and is 13.50. The invention further provides a beta-cyclodextrin inclusion compound PPHG / GA-beta-CD embedded with PPHG / GA, the synergic inhibition thrombin activity of PPHG / GA can be well preserved under the low-temperature and dark condition, it is shown that the beta-cyclodextrin inclusion compound PPHG / GA-beta-CD can better control and slowly release bioactive substances compared with monomeric substances, and the beta-cyclodextrin inclusion compound PPHG / GA-beta-CD can be applied to preparation of products for inhibiting thrombin activity and has good application prospects. Or the traditional Chinese medicine composition can be used for preparing products for treating thrombus diseases.

The invention discloses a thrombin activity detection kit and application thereof, and belongs to the technical field of biology. The unique characteristics of aggregation-induced emission (AIE) molecules are utilized, and the thrombin activity is monitored by detecting the change of luminophores in a blood coagulation system by virtue of the reaction process that fibrinogen is hydrolyzed by thrombin to generate fibrin monomers and the monomers are further self-polymerized into fibrin aggregates. Based on the principle, a thrombin activity detection method is established, a method for screening, separating and evaluating drugs containing potential thrombin inhibitor active components by taking immobilized thrombin as a medium is established, the thrombin inhibitor can be successfully screened out, and the inhibition effect of the thrombin inhibitor can be determined. A new method is provided for rapid detection of thrombin activity, and a new strategy is also provided for screening and evaluation of thrombin inhibitors.

The present invention relates to the technical field of rodent pest control. Disclosed are the use of avermectin and a rodenticide formulation. The avermectin has a structure represented by formula (I). The present invention uses a strategy of "new use for old drugs", uses commercially available pesticides as candidate compounds, and uses mouse thrombin as a target to single out pesticide avermectin strongly bound to thrombin, which can activate the activity of mouse thrombin and promote blood coagulation. The present invention has the advantages of low cost, good rodenticidal killing effect, acute and chronic efficacy, good palatability, easy degradation, and the like.

The application relates to the technical field of rodent control, and discloses application of an avermectin and a rodenticide preparation, wherein the avermectin has a structure shown in formula (I). The application uses the strategy of 'new use of old drugs', uses marketable pesticides as candidate compounds, and uses mouse thrombin as a target to screen the pesticide avermectin which can strongly bind to thrombin, can activate the activity of mouse thrombin, and can promote blood clotting; the application has the advantages of low cost, good rodenticide effect, combination of acute and chronic, good palatability, easy degradation and the like.

This invention discloses an antithrombin III activity detection reagent, comprising a heparin diluent, a thrombin reagent, and a chromogenic substrate reagent. The heparin diluent includes heparin, a first buffer solution, EDTA, sodium chloride, and a preservative. The thrombin reagent includes thrombin, a second buffer solution, a protective agent, an inorganic salt, and a preservative. The chromogenic substrate reagent includes a chromogenic substrate and a preservative. This invention uses a chromogenic substrate method, which offers high sensitivity and accuracy, improves the stability of the thrombin reagent, and ensures the reagent's safety and stability, while also demonstrating good synergistic effects among its components.

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.