17 results about "Heparinase" patented technology

The utility model provides a continuous flow micro-reactor for a heparinase catalytic reaction, belongs to the technical field of micro-reactors, and aims to solve the problems that the replacement of immobilized heparinase needs a long time and the efficiency of the heparinase catalytic reaction is influenced. Comprising a limiting supporting base, a positioning mounting frame, a reactor main body, a positioning connecting rod, a first storage tank and a second storage tank, the positioning mounting frame is in bolted connection with the right side of the limiting supporting base; the reactor main body is fixedly connected to the left side of the positioning mounting frame; the positioning connecting rod is rotationally connected to the left side of the reactor main body, and a damper is arranged at the joint of the positioning connecting rod and the reactor main body; the first storage tank is fixedly connected to the middle part of the positioning connecting rod; the second storage tank is in bolted connection with the front side of the first storage tank; according to the utility model, the replacement efficiency of the immobilized heparinase is improved, a large amount of time is saved, and the catalytic reaction efficiency of the heparinase is improved.

The present invention relates to an heparanase-binding and heparanase-neutralizing monoclonal antibody (IgG-1 mAb A54), including its epitope (HBD-II) and mode of interaction with heparanase, pharmaceutical composition comprising same, and uses thereof e.g. for inhibiting or treating a disease or disorder associated with heparanase activity, including but not limited to cancer, inflammation, viral infection, diabetes and related complications. The present invention further provides combinatorial cancer therapies, comprising the heparanase-neutralizing mAb and an additional anti-cancer treatment such as chemotherapy or radiation.

The invention belongs to the technical field of bioengineering, and relates to heparan sulfate defect type matrigel as well as a preparation method and application thereof. The preparation method comprises the following steps: providing natural matrigel; providing immobilized heparinase, wherein the immobilized heparinase can specifically catalyze and degrade heparan sulfate in the natural matrigel; carrying out contact reaction on the natural matrigel and the immobilized heparinase; and after the reaction is finished, separating the immobilized heparinase. According to the preparation method, the characteristics that the immobilized enzyme is convenient to separate and recycle are utilized, the problems that in a traditional solution enzyme method, enzyme residues exist, the reaction is difficult to accurately control, and subsequent purification steps are complex are solved, and a standardized heparan sulfate defective matrigel product which is controllable in component, free of residual enzyme and high in stability is provided. The method has wide application value in the fields of tumor mechanism research, anti-tumor drug screening, tissue engineering and the like.

The application discloses heparin oligosaccharide, a heparin oligosaccharide composition, a preparation method and application, and belongs to the technical field of biological medicines. The structure of the heparin oligosaccharide is as follows: heparin is enzymatically hydrolyzed into a low-molecular-weight heparin mixture; the low-molecular-weight heparin mixture is separated by a P10 chromatogram to obtain heparin decasaccharide; the heparin decasaccharide is separated by a strong anion exchange column to obtain salt-containing heparin oligosaccharide; and the salt-containing heparin oligosaccharide is desalted by a gel chromatogram column to obtain heparin oligosaccharide. The heparin oligosaccharide prepared by the application is beneficial to realizing quality control of a drug, improving the safety of the drug, and the heparin oligosaccharide is combined with SNAC, effectively promotes oral absorption of the heparin oligosaccharide, significantly improves the oral bioavailability of the heparin decasaccharide, has good anti-liver injury activity, and can be used for oral treatment of drug-induced acute liver injury.

The application belongs to the technical field of heparin extraction, and particularly relates to a method for preparing low-molecular heparan sulfate by using an enzyme method. The method comprises the following steps: (1) dissolving heparan sulfate in deionized water, adjusting the pH to 8-8.5 by using a NaOH solution at 20-25 DEG C, adding modified carbon nanotubes loaded with heparinase, then performing enzymolysis for 3.0-3.5 h under stirring at 35-45 DEG C, and obtaining an enzymolysis solution; (2) when the specific absorbance of the enzymolysis solution in step (1) is 1.3-1.5 at a wavelength of 230 nm, rapidly heating the enzymolysis solution to 95-98 DEG C, and terminating the reaction after 0.5-1.0 h; and (3) cooling the enzymolysis solution after the reaction to 20-30 DEG C, adding anhydrous ethanol, stirring for 1-2 h, centrifuging to obtain a precipitate, adding deionized water, filtering by using a 0.1-0.22 mu m filter membrane, and vacuum freeze-drying to obtain low-molecular heparan sulfate. The application has a mild reaction and a high yield.

The application belongs to the technical field of bioengineering, and relates to a heparan sulfate-deficient Matrigel as well as a preparation method and application thereof. The preparation method comprises the following steps: providing natural Matrigel; providing immobilized heparinase which can specifically catalyze degradation of heparan sulfate in the natural Matrigel; contacting and reacting the natural Matrigel with the immobilized heparinase; and separating the immobilized heparinase after the reaction is completed. The preparation method utilizes the characteristics of the immobilized enzyme that is convenient to separate and recover, overcomes problems such as enzyme residue, difficulty in accurately controlling the reaction and complexity of subsequent purification steps in the traditional solution enzyme method, and provides a standardized heparan sulfate-deficient Matrigel product which is controllable in composition, free of residual enzyme and high in stability. The application has wide application value in the fields of tumor mechanism research, anti-tumor drug screening and tissue engineering.

The present invention is directed to hexasaccharides mimetics of heparins presenting remarkable heparanase inhibition activity. The compounds have formula (I), wherein R1 is selected from the group consisting of NH2, NHSO3Na, NHAc; R2 is selected from the group consisting of NH2, NHSO3Na, NHAc; R3 is selected from the group consisting of Bn, H; R4 is selected from the group consisting of Ac, H, SO3Na; R5 is CO2Na; R6 is selected from the group consisting of Me, Ethyl, Alkyl, alkyl azide, alkynyl, cholestanol aglycon; R7 is selected from the group consisting of CH2OH, COOH.

The invention provides a construction method of a heparinase fingerprint spectrum, which comprises the following steps: a) taking a heparinase sample to be detected and diluting with a diluent to prepare a heparinase test solution with a certain concentration; and b) under normal pressure, determining the retention time and peak area A enzyme of each enzyme in the heparinase test sample solution by adopting a high performance liquid chromatography analysis method, and recording a spectrogram. The fingerprint spectrum of the heparinase and the construction method of the fingerprint spectrum can be widely applied to quality detection and control of a crude heparinase product and a finished heparinase product in the preparation process of the heparinase, and the method for detecting the quality of the heparinase by utilizing the fingerprint spectrum of the heparinase is simple, convenient, rapid and accurate to operate.

Embodiments of the present application disclose a heparinase detection method based on a magnetic bead method, applied to a sample analyzer, the method comprising: detecting a sample containing heparinase by using the magnetic bead method, determining a first coagulation time corresponding to the sample with heparinase according to collected first magnetic bead oscillation waveform data; as a comparison, detecting a sample without containing heparinase by using the magnetic bead method, determining a second coagulation time corresponding to the sample without heparinase according to collected second magnetic bead oscillation waveform data; and determining a detection result according to the first coagulation time and the second coagulation time. The method not only greatly reduces the cost, but also greatly improves the timeliness due to the simple detection method. In addition, a heparinase detection device based on the magnetic bead method, a sample analyzer and a storage medium are also proposed.

The application discloses a kind of composite lipid nanocapsule compositions and its preparation method and application, belong to medical technical field.The composite lipid nanocapsule composition is by the lipid core with negative charge on the surface, the first layer of capsule wall with positive charge, the second layer of capsule wall with negative charge, three are combined by electrostatic adsorption between positive and negative charge group Formed, can be used for intravenous injection.Lipid core is the depot for loading antitumor drug, chitosan has the effect of promoting transmembrane transport and controlling drug release, and the low molecular heparin coated outside nanocapsule can be reacted with heparinase, so that nanocapsule is enriched and degraded in malignant tumor site, realizes the function of targeting tumor.In addition, the carrier material used in the composition has high biological safety, good tumor targeting, high drug encapsulation efficiency, good stability, controllable release, and the preparation method is simple, and it has wide application prospect when used for the targeted delivery of antitumor drugs.

The invention provides a protein immobilization method and application in preparation of low-molecular heparin, a used modified carrier is prepared by infiltrating epoxy resin in an SST chimeric protein solution for combination, and one specific amino acid sequence of the SST chimeric protein is SEQ ID NO: 1; the nucleotide sequence of the coding gene is SEQ ID NO: 2. The invention also provides an immobilized enzyme, wherein the modified carrier is used as an immobilization medium to load protease; one amino acid sequence of the protease is SEQ ID NO: 5 or SEQ ID NO: 7. The protease is immobilized by adopting the modified ES-SST medium, so that the immobilized protease has remarkable advantages in the aspect of reutilization performance, the consumption of an enzyme preparation is reduced, and the production cost is reduced. After the prepared chimeric protein of the heparinase is immobilized, the catalytic characteristics of the heparinase, including enzyme activity, product distribution and optimal temperature and pH, can be more completely reserved, and the chimeric protein can be used for subsequent immobilization exploration.

The invention relates to marine-derived heparinase as well as a coding gene and application thereof, and belongs to the technical field of biological enzymes. The amino acid sequence of the heparinase HD1492 is as shown in SEQ ID NO. 1, and the nucleotide sequence of the heparinase HD1492 is as shown in SEQ ID NO. 2. The novel heparinase HD1492 mentioned in the invention is identified from a metagenome of marine bacteria cultured by taking HP as a unique carbon source, and shows unique enzymatic characteristics. The heparinase HD1492 preferentially cleaves the sulfated domain in the HP / HS chain, thereby producing a series of resistant oligosaccharide structures from tetrasaccharide to dodecasaccharide in the final digest of the low sulfated HS, and these oligosaccharides consist of non-sulfated disaccharides, or consist of at least one non-sulfated disaccharide linked at the reducing end to an N-sulfated disaccharide. Due to the characteristic, HD1492 is very useful in the aspect of researching distribution and composition of a non-sulfated structural domain in an HP / HS chain, and the limitation that only disaccharide composition information can be obtained through traditional enzyme analysis is overcome.

The invention discloses a cow leather preservation method for improving the extraction efficiency of collagen in cow leather, and belongs to the technical field of collagen preservation. The cattle hide with fat and muscle removed is placed in a mixed aqueous solution containing an enzymolysis regulator and a depilation enzyme preparation for enzymolysis depilation treatment, the cattle hide with depilation is placed in an antiseptic solution to be soaked, and cryopreservation is conducted at the cryopreservation temperature ranging from-5 DEG C to-20 DEG C after soaking; the enzymolysis regulator is prepared from mercaptoethanol, cysteine, calcium citrate and sodium dodecyl sulfate; the unhairing enzyme preparation consists of keratinase, heparinase, lipase and neutral amylase; the anti-corrosion solution is composed of glutaraldehyde, hydrochloric acid, zinc sulfate and water. When the method is adopted to cryopreserve the cow leather, the cryopreservation temperature can be reduced in the presence of the preservative solution, the loss of the cryopreserved collagen at the temperature is small, the cow leather can be preserved for about 6 months, and the application effect is good.

The invention belongs to the technical field of immune cell therapy, and particularly relates to a heparanase-loaded CAR-T exosome and application thereof. According to the invention, the HPSE-loaded exosome is secreted by the engineered CAR-T cell, and the activity of the HPSE is limited in a tumor microenvironment (TME) by utilizing an anchoring mechanism, so that systematic off-target toxicity is avoided. The result of the embodiment shows that the exosome shows remarkable ECM degradation capacity, tumor killing efficiency and immune regulation function in in-vitro and in-vivo models, has the characteristics of simplicity in preparation, high safety and remarkable curative effect, and provides a new solution for solid tumor immunotherapy.

The invention provides a large-scale multiplication culture method of CAR-T cells, and belongs to the technical field of CAR-T cells. The preparation method comprises the following steps: coupling bevacizumab and heparinase, immobilizing the coupled bevacizumab and heparinase on anti-CD3 / CD28 magnetic beads to prepare modified magnetic beads, resuspending CAR-T cells by using an MACS T cell culture medium containing potassium chloride, decitabine liposome and 2-deoxy-D-glucose, adding the modified magnetic beads, uniformly mixing, and performing magnetic field stimulation culture to obtain the high-activity CAR-T cells. The high-activity CAR-T cells prepared by adopting the large-scale amplification culture method of the CAR-T cells not only have better wettability, can enter a tumor microenvironment, but also can improve recruitment of the CAR-T cells in the tumor microenvironment, and have better anti-tumor activity, meanwhile, the amplification rate is obviously improved by adopting the method, and the method is suitable for large-scale in-vitro amplification culture.

The present invention is directed to synthetic hexasaccharide mimics of heparin having formula (I) wherein: R1 is selected from the group consisting of NH2, NHSO3 -, NHAc; R2 is selected from the group consisting of H, SO3 -, Bn, 2-Naphthylmethyl, preferably H; R3 is selected from the group consisting of H, Ac, SO3 -; R4 is CO2 -; R5 is Me, Et, C3-C30 linear or branched alkyl, optionally comprising one or more isolated or fused rings, PEG-OMe; R6 is selected from the group consisting of H, Ac, SO3 -; R is a C10-C30 linear or branched alkyl optionally comprising one or more isolated or fused rings. In another embodiment, the invention is directed to the use as a drug of hexasaccharides having the above formula, wherein R, R1, R2, R3, R4 and R6 have the above defined meaning, while R5 is selected from the group consisting of Me, Et, C3-C30 linear or branched alkyl, optionally comprising isolated or fused rings, PEG-OMe.