46 results about "Protein virus" patented technology

The invention relates to escherichia coli-derived single-plasmid-tandem soluble coexpression foot-and-mouth disease virus capsid proteins VP0 (which is a VP4 and VP2 fusion gene), VP1 and VP3, and a foot-and-mouth disease virus capsid protein virus-like particle preparation method. Foot-and-mouth disease virus capsid protein virus-like particles can be used for preparation of a foot-and-mouth disease vaccine. According to the method, a plurality of aspects of escherichia coli-derived soluble coexpression foot-and-mouth disease virus capsid protein are studied, by comprehensive use of tandem coexpression and SUMO(suggested upper merged ontology) technology with a tag for soluble coexpression of the foot-and-mouth disease virus capsid proteins VP0 (which is the VP4 and VP2 fusion gene), VP1 and VP3, the ultimate objective protein accounts for about 20% of total bacterial protein, and the foot-and-mouth disease virus capsid proteins obtained by purification can be successfully assembled into the virus like particles.

The invention discloses COVID-19-S-RBD virus-like particles and vaccine, and preparation methods of the virus-like particles and the vaccine. pET28a-CuMVTT recombinant plasmid is constructed after a CuMVTT gene is used to connect pET28a plasmid; pFUSE-COVID-19-S-RBD recombinant plasmid is constructed by a COVID-19-S-RBD gene and pFUSE plasmid; the recombinant plasmid is respectively transferred into the expression strains of escherichia coli and the expression cell lines of 293 F cells; the expression strains of escherichia coli are cultivated, biomass is separated through centrifugation, andthe virus-like particles are obtained; COVID-19-S-RBD protein is obtained by cultivating the expression cell lines of the 293 F cells; and the virus-like particles are coupled to the COVID-19-S-RBD through a chemical coupling reagent SMPH. The virus-like particles and the vaccine can be easily obtained through bacteria culture; yield can be higher than that of chimeric expression; and thus, industrial production and fast immunity can be achieved.

The invention discloses a method for preparing human blood coagulation factors IX and VII subcutaneously from cold-glue-removed blood plasma. The method comprises the following steps: 1, removing cold glue from blood plasma; 2, conducting strong anion-exchange column chromatography the first time; 3, conducting PEG sedimentation for removing impure protein; 4, conducting S/D viral inactivation; 5, conducting strong anion-exchange column chromatography the second time, and obtaining FVII eluent and FIX eluent; 6, conducting weak anion-exchange column chromatography, and concentration for purifying blood coagulation FVII; 7, conducting heparin affinity column chromatography for purifying blood coagulation FIX; 8, conducting ultrafiltration; 9, adding a stabilizing agent, and conducting adjustment; 10, conducting virus-removal filtration through nanofilms; 11, conducting sterilization, filtration and subpackage; 12, conducting freeze-drying; 13, conducting dry-hot viral inactivation. According to the method, PEG sedimentation is adopted for removing the impure protein, the target of preparing high-purity FVII and FIX simultaneously is achieved through combination of an ion-exchange column chromatography technology and an affinity chromatography technology, the process flow is simple, the production cycle is short, a product is subjected to three steps of virus eradicating measures, and use safety is high.

The invention discloses a monoclonal antibody BTV8-VP2-3E11 resistant to bluetongue virus 8 type (BTV8) VP2 protein, B-cell epitope peptide identified thereby and application, and belongs to the field of BTV8 control. A selected hybridoma cell strain capable of stably secreting a BTV8-VP2 protein resistant monoclonal antibody is stored with a microbial preservation number of CGMCC (China General Microbiological Culture Collection Center) No.7004. The experiment results show that the monoclonal antibody BTV8-VP2-3E11 secreted by the hybridoma cell strain can perform idiosyncratic reaction with the BTV8-VP2 protein but not reacts with the VP2 proteins of other serum types. The monoclonal antibody BTV8-VP2-3E11 and the BTV8-VP2 protein virus specific conserved B cell epitope peptide identified by the a monoclonal antibody can be prepared into an agent used for diagnosing BTV8 infection, thus laying a good foundation for creating serology differential diagnosis methods for BTV8 and other types of serum.

The present invention discloses a hybridoma cell line and an anti-canine distemper virus N protein monoclonal antibody CDV-1N8 produced through the hybridoma cell line, and belongs to the field of CDV prevention and treatment. According to the present invention, the hybridoma cell line stably secreting the anti-CDV N protein monoclonal antibody is screened, wherein the microorganism preservation number is CGMCC No.8793, and experiment results prove that the monoclonal antibody CDV-1N8 secreted by the hybridoma cell line can specifically react with the CDV N protein, and does not react with the Vero cell protein; the monoclonal antibody can specifically recognize the CDV-N protein, and accurate positioning of the CDV-N protein B-cell epitope recognized by the monoclonal antibody is QITFLHSERS; and the monoclonal antibody CDV-1N8 and the CDV N protein virus-specific conservative B cell epitope polypeptide recognized by the monoclonal antibody can be made into the CDV infection diagnosis reagent so as to establish the foundation for establishment of the CDV serological differential diagnosis method.

The invention provides a vaccine composition, which comprises an immunizing dose of PCV2Cap protein virus-like particle antigen and a stable buffer solution. In the vaccine composition, the PCV2Cap protein can stably exist in the form of virus-like particles, and after preservation of different time, the effect is obviously better than a contrast vaccine.

The invention discloses a monoclonal antibody BTV8-VP2-4D9 resistant to bluetongue virus 8 type (BTV8) VP2 protein, B-cell epitope peptide identified thereby and application, and belongs to the field of BTV8 control. A selected hybridoma cell strain capable of stably secreting a BTV8-VP2 protein resistant monoclonal antibody is stored with a microbial preservation number of CGMCC (China General Microbiological Culture Collection Center) No.7003. The experiment results show that the monoclonal antibody BTV8-VP2-4D9 secreted by the hybridoma cell strain can perform idiosyncratic reaction with the BTV8-VP2 protein but not reacts with the VP2 proteins of other serum types. The monoclonal antibody BTV8-VP2-4D9 and the BTV8-VP2 protein virus specific conserved B cell epitope peptide identified by the a monoclonal antibody can be prepared into an agent used for diagnosing BTV8 infection, thus laying a good foundation for creating serology differential diagnosis methods for BTV8 and other types of serum.

The invention discloses expression and purification of murine norovirus VP1 protein virus-like particles and preparation of a polyclonal antibody. According to the technical scheme, structural proteinVP1 genes of the murine norovirus are subjected to codon optimization for the first time, protein is expressed by utilizing a baculovirus expression system, the expressed protein forms the virus-likeparticles, and the antibody can simulate capsid protein of live viruses. For a VLPs antigen, the new Zealand white rabbit is immunized, the polyclonal antibody capable of resisting op.VP1 is prepared, the experiment proves that the polyclonal antibody can be used for Western Blot detection and IFA analysis, and a basic material is provided for detecting the MNV carrying condition of the laboratory mouse in a laboratory. Meanwhile, the VLPs antigen and the polyclonal antibody lay a foundation for further developing an ELISA detection kit.

The invention discloses a replication-defective A virus expression vector system and a vaccine preparation method. The vector system consists of a replication-defective A virus vector of a deleted structural gene and minus strand complementary plasmids or cells containing a controllable virus A structural gene. In the preparation method, an exogenous gene target is introduced by constructing the replication-defective A virus vector of a deleted virus structural protein; and the replication-recombinant A virus can be continuously produced in a large scale by combining the minus strand complementary plasmids or cells containing the controllable A virus structural gene, so that the problem of low virus yield of a conventional A virus system can be solved, a target of preparing recombinant virus in a large scale is realized, and exogenous protein and virus sample particles can be expressed in scale. The replication-defective A virus expression vector system disclosed by the invention is suitable for large-scale production of DNA (deoxyribonucleic acid) vaccine, vector virus vaccine and exogenous protein expression.

The invention supplies a manufacturing technique of IBV-HI antigen, the preparation process of antigen is that: the virus fluid of containing the protein virus is isolated from chicken embryo allantois fluid of containing respiratory type IBV the virus and blending with clostridium filtrate of rabbits A type wei surname as a culture medium, after two hours water bath action with 37deg.C, put the fluid into the 4deg.C fridge for 48 hours, through freeze drying to get antigen, the related centrifuge divided into two centrifugal, first the mixed fluid through high speed centrifugation, choose the supernatant after the centrifugation; then it progress the two centrifugation as ultracentrifugation, the time is 2-3h, the rotation rate of centrifugation is 25000-35000g, abandon the supernatant after the centrifugation, and use deposit suspension to obtain virus fluid. Using this invention method, infectious bronchitis HI antigen of respiratory system has relatively low cost, type specificity, high sensitivity, stability feature, it makes HI method to detect respiratory infectious bronchitis feasible, and enhance the credibility and maneuverability of its detection.

The present invention relates to the treatment of infectious diseases, specifically by extracorporeally eradicating the pathogen. This invention comprises methods for the extracorporeal treatment of infectious diseases that will remove infectious pathogens (leukemia cells, bacteria, viruses, or fungi causing a septicemia, metastatic cancer cells, target protein, viruses, parasites, fungi and prions) in humans by targeting such pathogens with a laser or other high-energy source of emissive radiation. More specifically, the method involves removing a bodily fluid from a patient, attaching an antibody to pathogens in the bodily fluid, sensing the antibody-pathogen moiety, using a high-powered, focused laser, or other suitable light source, to destroy the antibody-pathogen moiety, removing the remains of the antibody-pathogen by filtering or other suitable mechanism(s), and returning the bodily fluid to the patient.

The invention discloses a preparation method for porcine thrombin. The invention aims to solve the problems of high operation difficulty and being adverse to industrial production of the thrombin extraction. The key point of the technical scheme comprises the following steps: adding trisodium citrate liquid into pig blood, thereby acquiring a pig blood-trisodium citrate mixed liquid; centrifugallylayering the pig blood-trisodium citrate mixed liquid, thereby acquiring the plasma of pig blood; adding anion exchanger into the plasma of the pig blood, stirring and absorbing, cleaning the anion exchanger and then filling into an eluting column; preparing a NaCl-trisodium citrate mixed liquid as an eluant, eluting and collecting the eluant containing prothrombin; desalting the eluant, removingsolvent and monitoring the conductivity of the eluant eluted from a desalting column, thereby acquiring a desalted liquid containing prothrombin; adding CaCl2 liquid into an activated liquid containing thrombin; removing viruses, thereby acquiring a thrombin liquid. The impurities, such as, impure proteins and viruses in the thrombin can be effectively reduced and the thrombin purity can be increased.

A porcine circovirus type 2 (PCV2) Cap protein virus-like particle (VLP) preservation solution is based on a salt ion buffer solution system, saccharides, a nonionic surfactant, a metal ion chelatingagent, alcohols, a protein stabilizer, amino acids and a preservative are added, the PCV2 Cap protein VLP preservation solution can prolong the preservation period of the PCV2 Cap protein effectively,preserve the PVC2 Cap protein for 12 months at 2-8 DEG C while keeping protein concentration and activity unchanged, preserve the PCV2 Cap protein for 36 months at subzero 20 DEG C while keeping protein concentration and activity unchanged, can be used as a preservation solution of a standard protein of a kit for detecting the PCV antigen concentration or a diluted solution of a PCV2 Cap proteinsubunit vaccine.

The invention provides a canine parvovirus trivalence subunit vaccine. Antigens are VP2 protein virus-like particles of a canine parvovirus and are obtained by conducting cultivation and collection after a recombinant rhabdovirus is utilized for infecting insect cells; the recombinant rhabdovirus carries nucleotide fragments for coding VP2 proteins; the amino acid sequences of the VP2 proteins areSEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9 respectively. After the prepared vaccine is utilized for immunizing a canine, the antibody titer of the canine can be increased, and infection of the canine parvovirus is prevented.

A method for purifying a target protein from a cell culture sample containing the target protein, viral compounds and other impurities, by affinity chromatography virus inactivation and optionally other purifications the affinity chromatography involving

a) loading an affinity chromatography column with the cell culture sample thereby binding the target protein to the affinity chromatography column;

b) eluting the target protein from the affinity chromatography column by contacting the affinity chromatography column with an elution buffer having a pH<6 and comprising an excipient, wherein the excipient is disaccharides, polyols or poly (ethylene glycol) polymers;

c) collecting one or more fractions containing the target protein obtained from (b);

d) potentially combining the fractions obtained from (c) to form an elution product pool,

and wherein the virus inactivation involves

e) incubating the elution product pool at a pH from 2.5 to 4.5.

The invention discloses a medical nerve grafting repair film and a preparation method thereof. The preparation method comprises the steps of material selection, impure protein removal, virus inactivation, collagen degreasing, casing extension, chitosan atomization, dressing shaping, dressing solidification, cutting and inspection, packaging, sterilization and the like. Chitosan has antibacterial activity, can resist microbial infection, induce tissue regeneration and accelerate growth and healing of nerve fibers, and does not cause any inflammatory reaction in tissue; collagen has hemostatic and repairing functions, has bioabsorbability and cell adhesion, and can promote the formation of new cells and nerve cells; the product is low in antigenicity, good in human tissue compatibility, stable in absorption period through automatic coordination and absorption of enzymes and body fluid, and finally completely absorbed by a human body; the burn and scald repair film prepared by the methodhas the characteristics of uniform texture, good elasticity, high transparency, good air permeability, good biocompatibility, small toxic and side effects and the like, and the dressing can be absorbed by organisms.