31 results about "Prothrombinase" patented technology

The invention discloses a gymnadenia conopsea octasaccharide-decaose mixture as well as a preparation method and application thereof, and belongs to the technical field of medicines. The monosaccharide of the octasaccharide-decataccharide mixture of the gymnadenia conopsea comprises N-acetamino galactose, fucose, glucuronic acid or sugar alcohol thereof, galactose, N-acetylglucosamine, dehydrated talose and a sulfate group. The octasaccharide to decataccharide mixture of the gymnadenia conopsea disclosed by the invention can be used for remarkably prolonging the time of activating partial thromboplastin and prothrombin and effectively inhibiting endogenous and common coagulation pathways. The octasaccharide-decaose mixture of the gymnadenia conopsea can be used as a potential endogenous factor X enzyme inhibitor for preparing medicines for treating thrombotic cardiovascular diseases.

The invention relates to the technical field of detection kits, in particular to a blood coagulation reaction linked lupus anticoagulant fluorescence detection kit and application thereof, the kit contains a screening-confirmation integrated reagent, a calibration product, a quality control product and an anticoagulation interference inhibitor; the integrated reagent is divided into a reagent A and a reagent B, the reagent A contains a buffer solution, recombinant snake venom thrombin, composite phospholipid, a fluorescent substrate and other components, and the reagent B contains prothrombin, calcium ions, high-concentration composite phospholipid and other components; the composite phospholipid system is formed according to a specific proportion and modifies beta2-glycoprotein I, the fluorescence enhancement-stabilization system is formed by matching a Schiff base compound with a stabilizer, the calibrator is a series of lupus anticoagulant standard substances, and the quality control substance comprises simulants with different concentrations and negative control. The anti-interference capability is high, and the influence of anti-coagulant drugs and enzyme source fluctuation is reduced; the signal amplification effect is obvious, the detection sensitivity is high and the reagent stability is good; the operation is simple and convenient, screening and confirmation are integrally completed, and a conventional fluorescence detector is adapted.

The invention discloses a thrombin-antithrombin III compound determination method, a blocking antibody, a detection antibody and a kit, the blocking antibody blocks the vicinity of the 274-site residue of the prothrombin nitrogen terminal, and the detection antibody is used as a capture antibody of a TAT compound and specially aims at the effect of thrombin close to the thrombin N terminal. When the closing antibody near the position behind the 274-site residue of the prothrombin nitrogen terminal is combined with the prothrombin antibody, the closing antibody can effectively prevent the detection antibody at the N terminal of the thrombin from combining with the prothrombin, but because the closing antibody near the position behind the 274-site residue of the prothrombin nitrogen terminal cannot combine with the thrombin, the closing antibody can effectively prevent the detection antibody at the N terminal of the thrombin from combining with the prothrombin. Therefore, the combination of the detection antibody at the N end of the thrombin to the thrombin is not influenced, and the TAT compound detection process is not interfered by high-concentration prothrombin in the plasma.

We provide new monoclonal antibody inhibitors of coagulases, staphylocoagulase (“SC”) and von Willebrand factor-binding protein (“vWbp”) for treatment of S. aureus. The monoclonal antibodies are useful in targeting the SC N-terminus of SC and vWbp (respectively) and inhibiting prothrombin activation. The monoclonal antibodies are able to bind to and interfere with, modulate, and / or inhibit the binding interactions between the coagulase protein and its ligan protein prothrombin the activation of prothrombin.

The present invention relates to a method for producing thrombin from a thromboplastin source using a given BaSO4 reagent as an adsorbent for prothrombin, and a method for assessing the suitability of a given BaSO4 reagent for the preparation of thrombin. In at least one embodiment, the method comprises contacting a sample of the given BaSO4 reagent with a thromboplastin source to obtain BaSO4-adsorbed thromboplastin, and subsequently assessing the procoagulant activity of the BaSO4-adsorbed thromboplastin. In another embodiment, the method comprises the step of assessing the procoagulant activity relative to normal mammalian plasma.

The invention belongs to the technical field of polypeptide preparation and biological pharmacy, and particularly relates to eggshell membrane polypeptide with thrombin inhibition activity and application of the eggshell membrane polypeptide. According to the invention, two polypeptides are separated and purified from an eggshell membrane, and the two polypeptides are a nonapeptide YDGQHGRVS and an octapeptide EVPPSAL respectively. Tests prove that the nonapeptide YDGQHGRVS and the octapeptide EVPPSAL have thrombin inhibition activity equivalent to that of positive control heparin sodium, and discover that the nonapeptide YDGQHGRVS prolongs the activated partial thromboplastin time and the prothrombin time. The nonapeptide YDGQHGRVS and the octapeptide EVPPSAL provided by the invention are derived from eggshell membranes, are natural in source, good in thrombin inhibition activity and high in safety, and have a good application prospect.

The invention discloses a method for establishing a rat bionic model for sudden deafness caused by inner ear microcirculation disturbance, and relates to the technical field of animal model construction.The method comprises the steps that exosome extraction and purification are conducted, specifically, healthy rat bone marrow is obtained, mesenchymal stem cells are extracted and placed in a culture bottle to be cultured, cell culture supernate is collected, and the exosome is obtained; extracting exosomes by adopting an ultracentrifugation method; loading of the procoagulant factor and the exosome: selecting a prothrombin fragment as the procoagulant factor, treating the prothrombin fragment, and mixing the obtained exosome resuspension and the prothrombin fragment solution according to the volume ratio of 1: 1. The exosome derived from the mesenchymal stem cells is adopted as a procoagulant factor carrier, the source is wide, and the exosome is easy to obtain; the exosome is stable in yield and good in biocompatibility, damage to inner ear tissues of rats can be reduced, and the exosome has natural targeting transportation capacity and can be specifically combined with inner ear vascular endothelial cells, so that the concentration of a procoagulant factor at a target site is increased, and the accuracy of model establishment is enhanced.

The application provides an abnormal prothrombin detection kit and application thereof, and relates to the technical field of in-vitro detection.The application provides a kit for abnormal prothrombin detection, which comprises an abnormal prothrombin antibody, wherein the abnormal prothrombin antibody is derived from a hybridoma cell strain 7E8-4-4-1 of fusion of myeloma cells and mouse B cells, and the hybridoma cell strain 7E8-4-4-1 is preserved in the China General Microbiological Culture Collection Center on October 28, 2025, with a preservation number of CGMCC No.46709.The kit has a large detection range, a small lower limit of detection and high accuracy, and has great application potential in the detection of abnormal prothrombin.

The present invention discloses a method for simultaneously separating canine fibrinogen and canine prothrombin complex from canine plasma. First, rough separation and fine separation are carried out by gel column chromatography and anion column chromatography. Then, further purification is carried out by DEAE Sephadex A-50 gel adsorption. Through the combination of multiple steps of processes and their parameters, high-purity and high-yield canine fibrinogen, prothrombin and other components can be extracted from canine plasma simultaneously. The purity of both canine fibrinogen and canine prothrombin complex can reach over 95%, and the extraction rates of canine fibrinogen and canine prothrombin complex reach over 90% and over 85% respectively, greatly improving the comprehensive utilization rate of canine plasma. At the same time, by adjusting the formula and ratio of the stabilizer, the freeze-dried products of canine fibrinogen and canine prothrombin complex have good solubility, clear solutions, and short reconstitution times, which are 10-15 minutes and 5-10 minutes respectively. In summary, the method of the present invention has stable processes, high raw material utilization rate, and good product quality.

The invention relates to a composition for stabilizing a human prothrombin complex and application of the composition. The composition comprises an anticoagulant and a protective agent, the anticoagulant is heparin and / or a pharmaceutically acceptable salt thereof; the protective agent is selected from arginine and / or histidine and one or more of their respective pharmaceutically acceptable salts, and the ratio of the anticoagulant to the protective agent is (4.5-9.5) IU: (0.02-0.04) g. The activity of FII, FVII, FIX and FX of the prothrombin complex preparation prepared by adopting the composition for stabilizing the human prothrombin complex provided by the invention is not obviously reduced even under the condition of high temperature (40 + / -2 DEG C), and the activity of the FII, the FVII, the FIX and the FX is not obviously reduced after the prothrombin complex preparation is stored at the high temperature for at least 24 months. The biological activity index of the prothrombin complex preparation still meets the clinical use standard.

Overt hepatic encephalopathy is a serious complication of cirrhosis and affects lipid and lipoprotein metabolism. The present invention relates to a biomarker for the prognosis of hepatic encephalopathy. The biomarker includes one or more of high-density lipoprotein cholesterol, model for end-stage liver disease score, age, gender, upper gastrointestinal bleeding, spontaneous bacterial peritonitis, overt hepatic encephalopathy grade, ascites, aspartate aminotransferase, alanine aminotransferase, total bilirubin, serum albumin, total cholesterol, low-density lipoprotein cholesterol, serum creatinine, neutrophil-lymphocyte ratio, prothrombin time, and prothrombin activity level. Compared with patients with alcoholic hepatitis, liver disease, liver failure, etc. proposed in the prior art, due to the specificity of the target population, the biomarker proposed in this application can significantly improve the accuracy of the assessment of the prognosis of overt hepatic encephalopathy.

The utility model discloses an auxiliary experimental device for determining the content of heparin in a human prothrombin complex, and belongs to the technical field of experimental devices. The device comprises a vibrator, a test tube rack and an illuminating lamp B. The test tube rack is arranged on the vibrator, and the illuminating lamp B is located over the test tube rack and connected with the vibrator through a support. A reaction reagent and a sample to be detected in the test tube are automatically shaken by adopting the vibrator, so that both hands of an experimenter are liberated, and the experimenter can conveniently observe the solidification condition of liquid in the test tube; the lighting lamp B is used for supplementing light, so that an experimenter can quickly and accurately observe the solidification condition of the liquid in the test tube.

The invention relates to the technical field of biological medicine, and provides a plasma metabolite-based ALC and AH noninvasive differential diagnosis model, which is characterized in that five plasma key metabolites are taken as core indexes, and a probability calculation model is constructed through a logistic regression algorithm; the five kinds of plasma key metabolites are respectively an Ab-fubinaca metabolite 3, isocitric acid, N-docosahexaenoyl cysteine, gluconic acid-delta-lactone, and 1-palmitoyl-2-linoleyl phosphatidylethanolamine. The invention further provides a construction method and application of the model. According to the method, five key plasma metabolites with remarkable difference between AH and ALC are screened, modeling is carried out in combination with a logistic regression algorithm, or a multi-parameter model is constructed by further integrating prothrombin time, so that accurate non-invasive efficient identification of AH and ALC is realized.

The invention discloses an abnormal prothrombin antibody and application thereof, and relates to the field of antibodies. The abnormal prothrombin antibody disclosed by the invention comprises a heavy chain complementarity determining region and a light chain complementarity determining region, provides an important raw material source for detection of abnormal prothrombin, and has good affinity or activity.

The invention provides a prothrombin activation process and a thrombin preparation method, according to the prothrombin activation process, a phospholipid substance is adopted as an activating agent, a calcium chloride solution is used for providing a calcium ion environment, and polysorbate 80 is adopted for assisting dissolution, so that prothrombin generates thrombin, and the invention further discloses the thrombin preparation method. Comprising the following steps: 1) collecting pig blood; 2) centrifugal separation; 3) adsorption separation; (4) eluting prothrombin; (5) carrying out prothrombin activation by adopting the prothrombin activation process; 6) ultrafiltration concentration; according to the prothrombin activation process provided by the invention, phospholipid substances are matched with polysorbate 80 and calcium chloride to serve as thrombin activators, the polysorbate 80 contributes to dissolution of the phospholipid substances, and compared with rabbit brain powder, the method has the advantages that the safety of the product can be remarkably improved, and compared with soybean phospholipid or egg yolk phospholipid, the stability of the product is greatly improved. The prothrombin activation rate can be obviously improved, and the method is suitable for large-scale production.

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 dominant hepatic encephalopathy is a serious complication of liver cirrhosis and affects lipid and lipoprotein metabolism. The invention relates to a biomarker for prognosis of hepatic encephalopathy. The biomarkers comprise high-density lipoprotein cholesterol, an end-stage liver disease model score, age, gender, upper gastrointestinal hemorrhage, spontaneous bacterial peritonitis, dominant hepatic encephalopathy grading, ascites, aspartic transaminase, glutamic-pyruvic transaminase, total bilirubin, serum albumin, total cholesterol, low-density lipoprotein cholesterol and serum creatinine. The specific value of neutrophil to lymphocyte, the prothrombin time and the prothrombin activity level are one or more. Compared with patients with alcoholic hepatitis, liver diseases, hepatic failure and the like in the prior art, the marker provided by the invention can remarkably improve the evaluation accuracy of the prognosis of the dominant hepatic encephalopathy due to the specificity of people.

The invention relates to an anti-prothrombin / phosphatidylserine antibody, a pharmaceutical composition and an application of the anti-prothrombin / phosphatidylserine antibody. The anti-prothrombin / phosphatidylserine antibody provided by the invention is connected with a prothrombin / phosphatidylserine compound with high affinity, so that the antibody is combined with phosphatidylserine in a targeted manner to achieve the treatment of tumor resistance, infection resistance, thrombosis resistance, atherosclerosis, ischemia-reperfusion injury, inflammatory bowel disease, Alzheimer's disease and Parkinson's disease.

The present invention discloses a prothrombin time detection reagent and a preparation method thereof. Among them, the prothrombin time detection reagent is prepared from rabbit brain powder, a solvent, sodium chloride, a surfactant, a metal salt, a reducing agent, and calcium chloride. It can effectively remove coagulation factor VII, so that the prepared thromboplastin does not contain coagulation factor VII, making the prepared prothrombin time detection reagent highly sensitive to coagulation factor VII, ensuring the detection accuracy and improving its actual use effect.

The present invention relates to the field of medical detection, and specifically to a portable device for prothrombin time detection. The prothrombin time detection device includes a detection area fixed with aggregation-induced luminescence carbon dots, and the detection area is used to receive plasma; the aggregation-induced luminescence carbon dots are synthesized by the following method: dithiosalicylic acid or citric acid is mixed with triethylenetetramine or tetraethylenepentamine, and then obtained by reaction. The carbon dots of this scheme have good biocompatibility and viscosity response characteristics. The plasma sample activates the exogenous coagulation pathway under the action of coagulation reagents. Under the action of continuous excitation light, the aggregation-induced luminescence carbon dots generate a fluorescence-time curve, and its emitted light produces a red shift phenomenon, thereby determining and calculating the prothrombin time. This technical scheme can solve the technical problems of the prothrombin time detection device of the prior art that the detection accuracy and stability are not ideal due to its reliance on electrochemical detection methods, and has an ideal application and promotion prospect.

The present invention belongs to the field of biotechnology engineering and provides a recombinant tissue factor (rhTF) mutant and its application. The recombinant tissue factor mutant is: rhTF mutant A, whose amino acid sequence is shown in SEQ ID NO.3 and whose mutation sites are simultaneously mutated: T17F, I22R, S47R, T112W, S188R, and V207R, and whose nucleotide sequence is shown in SEQ ID NO.4; or rhTF mutant B, whose amino acid sequence is shown in SEQ ID NO.5 and whose mutation sites are simultaneously mutated: T17F, I22R, S47F, T112W, S188K, and V207K, and whose nucleotide sequence is shown in SEQ ID NO.6. These mutants have high procoagulant activity and good thermal stability and are used in biomedicine, clinical prothrombin time determination, and wound hemostasis.

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.

To provide an antibody for factor xa inhibitor and method for using the same.SOLUTION: One of the major limitations of anticoagulant therapy is the limitation of the ability to rapidly reverse anticoagulant activity in the case of treatment-related bleeding risk and in cases of overdosage or required emergency surgical intervention. Therefore, a specific effective antidote for all forms of anticoagulant therapy is highly desirable. The present invention relates to an antidote to an anticoagulant that targets factor Xa. The antidotes are 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 described herein lack or have reduced intrinsic coagulant activity. Disclosed herein are methods of stopping or preventing bleeding in a patient that is currently undergoing anticoagulant therapy with a factor Xa inhibitor.SELECTED DRAWING: None

The invention relates to the technical field of antibodies, in particular to an abnormal prothrombin (PIVKA-II) antibody or a conjugate thereof and application thereof. The antibody or the conjugate thereof comprises a heavy chain variable region VH and a light chain variable region VL, the amino acid sequences of HCDR1, HCDR2 and HCDR3 of the heavy chain variable region VH are respectively shown as SEQ ID No: 1-3, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 of the light chain variable region VL are respectively shown as SEQ ID No: 4-5; the antibody conjugate is formed by coupling the antibody with a marker or a solid-phase carrier. The antibody disclosed by the invention has high binding activity and affinity to PIVKA-II, and the specificity, sensitivity, reliability and accuracy detection of trace PIVKA-II in a sample can be realized in a double-antibody sandwich manner.

Provided in the present invention is an anti-prothrombin / phosphatidylserine antibody, comprising a heavy chain variable region and a light chain variable region; the heavy chain variable region comprises HCDR1 to HCDR3 having amino acid sequences as shown in SEQ ID NOs: 1-3, respectively, and the light chain variable region comprises LCDR1 to LCDR3 having amino acid sequences as shown in SEQ ID NOs: 4-6, respectively. The anti-prothrombin / phosphatidylserine antibody of the present invention can bind to a prothrombin / phosphatidylserine complex with high affinity, thereby targeting and binding to phosphatidylserine, and has a potential therapeutic effect with respect to tumors, infections, atherosclerosis, ischemia-reperfusion injury, inflammatory bowel disease, Alzheimer's disease, Parkinson's disease, etc.

The invention belongs to the technical field of biology, and particularly relates to an anti-human thrombin-antithrombin complex antibody and a preparation method and application thereof and a preparation method of an anti-human thrombin-antithrombin complex (TAT), and the preparation method is characterized by comprising the following steps: (1) expressing and purifying human prothrombin (PT) and human antithrombin III (AT-III); (2) activating PT by using Ecarin snake venom to obtain thrombin (T); and (3) mixing T and AT-III according to a mass ratio of 0.7: 1 for reaction, and purifying to obtain the TAT complex, the invention also comprises a monoclonal antibody secreted by a hybridoma cell strain TAT-2F for specific recognition of TAT and a monoclonal antibody secreted by a hybridoma cell strain T-5B for specific recognition of T and TAT, and the monoclonal antibody provided by the invention can be combined with TAT with high specificity, so that the TAT complex can be used for detecting TAT and TAT. Therefore, the monoclonal antibody can be used for identifying or detecting a patient with abnormal blood coagulation.

The application provides a camel milk casein anticoagulant peptide, a preparation method and application, and the camel milk casein anticoagulant peptide is ISQFYQKWK. The application takes camel milk casein as raw material, obtains the target anticoagulant peptide through enzymolysis, separation and identification and solid-phase synthesis, solves the problems that existing anticoagulant drugs have bleeding risk, are relatively high in cost and are limited in long-term application, and simultaneously improves the high-value utilization level of camel milk casein resources. The camel milk casein anticoagulant peptide is natural in origin, clear in sequence, has good in-vitro anticoagulant activity, can intervene in the blood clotting process by prolonging the activated partial thromboplastin time, the prothrombin time and the thrombin time.

The present disclosure relates to antidotes to anticoagulants targeting factor Xa. The antidotes are factor Xa protein derivatives that bind to the factor Xa inhibitors thereby substantially neutralizing them but do not assemble into the prothrombinase complex. In one embodiment, the derivatives described herein lack or have reduced intrinsic coagulant activity.

The invention provides a prothrombin enzyme compound. The prothrombin enzyme compound comprises washed human plasma platelets, a human blood coagulation factor FXa and human prothrombin, or comprises Platelin LS, a human blood coagulation factor FXa, a human blood coagulation factor FVa and a human prothrombin, or comprises Platelin LS, a human blood coagulation factor FXa and a human prothrombin. The prothrombinase compound provided by the invention can monitor the activity of FV and FXa, can be used for confirming an AiFVD case and monitoring the hemostasis condition of a patient after treatment; or for monitoring the effectiveness of the platelet infusion product; or the method is used for screening the anticoagulant drugs from the candidates.

The invention discloses a method for purifying and activating prothrombin, which comprises the following steps: by taking cryoprecipitate-removed plasma as a raw material, washing and eluting by adopting an anion exchange resin adsorption method to obtain a solution containing the prothrombin and a blood coagulation factor X (FX), carrying out ultrafiltration on the solution to reduce the conductivity to be below 2.5 ms / cm, and concentrating the concentration of the prothrombin to be above 15IU / ml to obtain the prothrombin. And adjusting the concentration of FX to 50%-150% of the concentration of prothrombin. Adjusting the concentration of the prothrombin, adding a calcium chloride solution into the prothrombin until the final concentration of calcium chloride in the solution is 20-60mmol / l, uniformly stirring and mixing, and activating for 6-15 days at the temperature of 0-8 DEG C to obtain a thrombin solution. According to the method, the prothrombin recovery rate is high, the activation effect is good, and each IU of prothrombin can be activated to obtain 207.5 IU of thrombin titer. The problem that the thrombin titer recovery rate is not high is solved, and the yield of unit plasma thrombin titer is greatly increased.