141 results about "Virus antigen" patented technology

The invention discloses a porcine epidemic diarrhea virus antigen fusion protein, a coding gene and a porcine epidemic diarrhea vaccine prepared from the porcine epidemic diarrhea virus antigen fusion protein. The method comprises the following steps: screening to obtain a homosensory sequence of the spike protein of the porcine epidemic diarrhea virus, and fusing the homosensory sequence with a monomeric porcine ferritin heavy chain subunit to obtain an antigen fusion protein; in order to improve the expression quantity or the titer of the antigen fusion protein, the obtained antigen fusion protein is subjected to sequence optimization and then subjected to single-site or multi-site mutation, and the expression quantity and the titer of the obtained mutant are remarkably improved. According to the invention, a recombinant protein is expressed by using a silkworm or AcMNPV-insect cell and other eukaryotic expression systems to obtain a self-assembled ferritin nanoparticle antigen for displaying an S protein antigen protein on the surface of a cage structure of porcine ferritin, and a vaccine prepared from the prepared nanoparticle antigen can induce a widely neutralized anti-porcine epidemic diarrhea virus antibody; the porcine epidemic diarrhea virus vaccine has the potential of becoming a universal porcine epidemic diarrhea virus vaccine with cross immune efficacy.

The present invention discloses a porcine Seneca virus neutralization liquid phase blocking ELISA kit, which comprises an effective amount of porcine Seneca virus antigen, an effective amount of porcine Seneca virus single domain antibody 1, an effective amount of porcine Seneca virus single domain antibody 2, and a matching detection reagent. The porcine Seneca virus antigen is porcine Seneca virus particles purified by sucrose density gradient centrifugation, and the particle size of the purified porcine Seneca virus particles is 20 to 30 nm. The amino acid sequence of the heavy chain variable region of the porcine Seneca virus single domain antibody 1 is shown in SEQ ID NO.1. The amino acid sequence of the heavy chain variable region of the porcine Seneca virus single domain antibody 2 is shown in SEQ ID NO.2. The kit of the present invention has the advantages of high efficiency, simplicity, sensitivity, stability, etc., has a good correlation with the traditional diagnostic method virus neutralization experiment, and is suitable for a wide range of applications.

The invention provides a respiratory syncytial virus antigenic polypeptide, nucleic acid and a vaccine. Compared with the respiratory syncytial virus wild type Pre-F protein, the respiratory syncytial virus antigenic polypeptide or the immunogenic fragment of the respiratory syncytial virus antigenic polypeptide has the following changes: the 104th to 144th regions of the Pre-F protein are replaced with GS linker; two groups of disulfide bonds are introduced into S155C, S290C, A149C and Y458C; the S190F and the V207L are used for realizing cavity filling; and P102A, L373R, I379V, M447V, P102A, L373R, the C terminal of the respiratory syncytial virus antigenic polypeptide or the immunogenic fragment thereof comprises a cytoplasm tail region fragment, and the cytoplasm tail region fragment is a fragment derived from 3-10 continuous amino acid residues of a PreF protein cytoplasm tail region. According to the present invention, the pre-F immunogenicity and / or stability can be improved, and the protein expression level can be easily improved.

Provided are a respiratory syncytial virus antigenic polypeptide, a vaccine, and a use. First, a respiratory syncytial virus antigenic polypeptide or an immunogenic fragment thereof is provided. Compared with the amino acid sequence of a respiratory syncytial virus wild-type pre-F protein, the amino acid sequence of the respiratory syncytial virus antigenic polypeptide or the immunogenic fragment thereof has one or more mutation sites of amino acid residues at the following positions: positions 174-176, positions 181-184, positions 210-213, and positions 160-161. The improvement of pre-F immunogenicity can be achieved and / or the protein expression level can be improved.

The utility model relates to the field of antigen detection kits, and discloses an infectious bovine rhinotracheitis virus antigen detection kit which comprises a box body, a roller is rotatably connected to the bottom side in the box body, a clamping groove is formed in the upper side of the roller, a clamping pin is connected to the clamping groove in a clamping mode, and a placing table is fixedly connected to the upper side of the clamping pin. A plurality of swab placing grooves, an extraction tube placing groove and a detection card placing groove are respectively formed in the upper side of the placing table, the plurality of swab placing grooves, the extraction tube placing groove and the detection card placing groove are distributed in a circumferential array, and sampling swabs are arranged on the inner sides of the plurality of swab placing grooves; according to the utility model, a plurality of groups of sampling swabs, sample extraction tubes and antigen detection cards which are distributed in a circumferential array manner are placed on the placement table, so that when sampling personnel go to cattle for sampling one by one, the sampling personnel can rotate to detection materials such as the next group of antigen detection cards and the like only by pressing down the push rod and switching the angle of the placement table, and the operation is convenient and the use is simple.

The invention discloses a preparation method of a triple detection kit for influenza A, influenza B and new coronavirus antigens, and belongs to the field of respiratory virus in-vitro detection. In the antibody labeling stage, the antigen and the antibody can be more sufficiently combined in a manner of adding the linking arm, so that the antigen detection sensitivity can be improved. In the metal spraying stage, a novel coronavirus monoclonal antibody, an influenza A virus monoclonal antibody and an influenza B virus monoclonal antibody marked by latex microspheres are sequentially subjected to multi-layer metal spraying at the same position by a metal spraying instrument, the two antibodies are isolated by a trehalose solution, and metal spraying of three layers of antibodies isolated from each other is completed. And the cross reaction between different antibodies of the conjugate pad in the storage process is avoided, so that non-specific binding is avoided, and the storage stability is improved. The detection kit prepared by the method can specifically detect the novel coronavirus, the influenza A virus and the influenza B virus, and has the advantages of high sensitivity, good repeatability and the like.

The invention relates to the technical field of immunodetection, and discloses a bovine nodular skin disease virus chimeric antigen and application thereof in an ELISA (enzyme-linked immuno sorbent assay) detection kit. According to the chimeric antigen, a gene of a dominant region of a bovine nodular skin disease virus antigen is selected to design a chimeric antigen gene, meanwhile, sequence optimization is performed, a chemical synthesis method is used for synthesizing a coding gene of the chimeric antigen of the bovine nodular skin disease virus, and then the chimeric antigen is obtained in a prokaryotic expression mode. An ELISA antibody detection kit developed on the basis of the chimeric antigen has extremely high sensitivity and specificity, can rapidly and specifically detect bovine nodular skin disease virus antibodies in serum, is suitable for monitoring and checking the nodular skin diseases of cattle in large-scale breeding, and has relatively high practical value.

The invention relates to the technical field of genetic engineering, in particular to Encapsulin fusion protein for expressing foot and mouth disease virus epitopes and application of the Encapsulin fusion protein in preparation of subunit vaccines. The invention discovers that different serotypes of foot-and-mouth disease virus antigen epitopes are inserted among the 62nd to 63rd positions, the 124 to 125th positions, the 138 to 139th positions and the 239th to 240th positions of amino acids of an Encapsulin protein fragment subjected to amino acid sequence modification, and / or the 62nd to 63rd positions, the 124 to 125th positions, the 138 to 139th positions and the 239th to 240th positions of amino acids are replaced by the different serotypes of foot-and-mouth disease virus antigen epitopes; efficient and soluble expression of a target antigen in escherichia coli can be realized, and protein cage nano antigen particles are successfully self-assembled; the protein nano antigen particle can induce a widely neutralized foot-and-mouth disease virus antibody, improves the immune efficacy, and has the potential of becoming a broad-spectrum multivalent vaccine for foot-and-mouth disease.

The present invention relates to a kit for detecting porcine epidemic diarrhea and a preparation method thereof, belonging to the technical field of virus detection, and specifically comprising the following steps: after extracting the antigen, treating it with a virus purification agent containing polyethylene glycol, nuclease and concanavalin, and then obtaining a virus antigen suspension with a composite dissociation solution. The suspension is coated on a microporous reaction plate to form a solid-phase antigen. Subsequently, the porcine epidemic diarrhea virus solid-phase antigen, enzyme marker, sample diluent, Tris buffer, color developer and stop solution are aseptically stored to prepare a kit for detecting porcine epidemic diarrhea. The present invention provides a novel kit and a preparation method thereof, which can not only effectively purify porcine epidemic diarrhea virus, but also significantly reduce the risk of protein precipitation, while overcoming nucleic acid interference and improving purification efficiency.

The present invention relates to the field of biomedicine, and particularly to a T cell receptor and its preparation method and use. The T cell receptor EBV-TCR of the present invention comprises a variable region of the TCRα chain and a variable region of the TCRβ chain; the variable region of the TCRα chain contains peptide segments CDRα1, CDRα2, and CDRα3; the variable region of the TCRβ chain contains peptide segments CDRβ1, CDRβ2, and CDRβ3; the amino acid sequence of CDRα3 is as shown in SEQ ID NO.7, and the amino acid sequence of CDRβ3 is as shown in SEQ ID NO.3. The TCR of the present invention has high specificity, and binding to T cells can significantly activate cytokine release, providing a solid theoretical and experimental basis for the development of TCR-T therapy targeting Epstein-Barr virus antigens.

The invention relates to methods and kits for detecting a virus, e.g., a respiratory virus such as a coronavirus, in a biological sample. The invention also relates to methods and kits for detecting and / or quantifying biomarkers, e.g., antibody biomarkers against a viral antigen; inflammatory and / or tissue damage response biomarkers; and / or extracellular vesicles in response to a viral infection.

The invention provides a respiratory syncytial virus antigenic polypeptide, nucleic acid, a vaccine and application thereof. The invention firstly provides a respiratory syncytial virus antigenic polypeptide or an immunogenic fragment thereof, and the respiratory syncytial virus antigenic polypeptide has at least 26th-470th or full-length amino acid sequence as shown in SEQ ID NO: 1, or a derivative amino acid sequence which is formed by replacing, deleting or adding one or more amino acids on the basis of the amino acid sequences and has the same function. According to the invention, the pre-F immunogenicity can be improved and / or the protein expression level can be improved.

The invention relates to the technical field of virus antigens, and particularly discloses an inactivation method for improving the recovery rate of enterovirus antigens. The method disclosed by the invention specifically comprises the following steps: adding a protective agent with the final concentration of 50-250 [mu] g / ml into purified enterovirus liquid; after fully and uniformly mixing, stirring and incubating the mixed solution; the protective agent is prepared by mixing TritionX-100, arginine and lysine; the incubation temperature is 25-40 DEG C, and the incubation time is 4-12 hours; then adding a formaldehyde solution until the final concentration is 180-220 mu g / ml, and inactivating for 68-76 hours under the condition that the temperature is 35-39 DEG C, so as to obtain an inactivated vaccine; and finally, washing and filtering the inactivated solution to remove TritionX-100 and formaldehyde in the sample. According to the technical scheme, the recovery rate of the antigen in the inactivation solution is greatly improved, and the residual quantity of TritionX-100 and formaldehyde is small.

The invention relates to a linear covalent closed DNA vaccine, which comprises a plasmid vector containing an IHNV virus antigen expression unit, the sequence of the antigen expression unit is shown as SEQ ID NO.1, and the DNA structural form is a linear terminal covalent closed form. The invention also relates to a preparation method and application of the linear covalent closed DNA vaccine. Compared with a traditional DNA vaccine, the linear covalent closed DNA vaccine has no redundant bacterium source nucleic acid sequence, and has good safety and effectiveness for preventing and treating infectious hematopoietic necrosis of rainbow trout.

The invention relates to the technical field of genetic engineering, in particular to a self-assembled protein cage nanoparticle epitope insertion site and application thereof in vaccine preparation. It is found that a foot-and-mouth disease virus antigen epitope is inserted between the 124 -125 amino acid of an Encapsulin protein fragment subjected to amino acid sequence modification, and / or the 124 -125 amino acid is replaced with the foot-and-mouth disease virus antigen epitope, efficient and soluble expression of a target antigen in escherichia coli can be achieved, and protein cage nano antigen particles are successfully self-assembled; the protein nano antigen particle can induce a widely neutralized anti-foot-and-mouth disease virus antibody, improves the immune efficacy, and has the potential of becoming a foot-and-mouth disease vaccine.

The invention discloses an EB (Epstein-Barr) virus antigen with chimeric design and application thereof. The EB virus chimeric antigen provided by the invention is reasonably designed on the basis of structural biology, gL, gH and gp42 proteins of the EB virus are connected through a linker, and the chimeric designed antigen stably presents antigen epitopes of the three proteins at the same time, so that the production amplification process and quality control can be simplified while the vaccine immunogenicity is improved. The EB virus chimeric antigen can be used independently or in combination with other antigens, has high immunogenicity when being used as a vaccine or a vaccine component, can induce an immunized animal to generate a high-level neutralizing antibody, can be used for preparing a vaccine for preventing or treating EB virus infection, and can also be used as a reagent for detecting EB virus.

The present invention discloses a neutralizing monoclonal antibody against Epstein-Barr virus gB antigen and its application. The antibody comprises a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises heavy chain CDR-H1 shown in SEQ ID NO:15; heavy chain CDR-H2 shown in SEQ ID NO:16; and heavy chain CDR-H3 shown in SEQ ID NO:17. The light chain variable region comprises light chain CDR-L1 shown in SEQ ID NO:18; light chain CDR-L2 shown in SEQ ID NO:19; and light chain CDR-L3 shown in SEQ ID NO:20. The antibody in the present invention can specifically recognize Epstein-Barr virus gB antigen and has a good neutralizing effect, so that various expression vectors and transformants can be developed based on the antibody, and thus can be effectively used for the detection, prevention and treatment of Epstein-Barr virus.

The present disclosure provides multicistronic vaccines and method for producing and using the same in preventing infection or transmission, or reducing severity of disease caused by influenza and / or SARS-Cov-2 virus in a subject. Multicistronic vaccines of the disclosure can be administered via intramuscular, intranasal, or inhalation route. In one particular embodiments, the disclosure provides a recombinant adenovirus comprising at least two different extraneous oligonucleotides that are capable of stimulating an immune response in a subject. Each of the oligonucleotide independently comprises an oligonucleotide that encodes either influenza or SARS-Cov-2 virus antigens.

The invention relates to a detection kit for nasopharynx cancer diagnosis and screening, which comprises an immunochromatography detection card, and the immunochromatography detection card comprises a reagent for detecting EB virus antigen and antibody through a colloidal gold labeling method. The invention also relates to application of the detection card and the EB virus antigen-antibody in preparation of a detection kit or a detection card for nasopharynx cancer diagnosis and screening.

The present invention relates to the field of biomedicine technology, and specifically discloses an mRNA based on the CP protein gene of fish nervous necrosis virus, a vaccine, and a preparation method and application thereof. The present invention provides an mRNA of fish nervous necrosis virus, the nucleotide sequence of which is shown in SEQ ID NO: 1, and the structure includes a 5' untranslated region, a signal peptide sequence, an NNV virus antigen coding region, a 3' untranslated region and polyA. The mRNA vaccine contains the ORF sequence of the capsid protein CP gene that is resistant to NNV virus infection. The results of animal safety tests on the fish nervous necrosis virus mRNA vaccine prepared by the present invention show that the vaccine is safe and effective, and is convenient and simple to use clinically, and has broad application prospects in the prevention and control of viral nervous necrosis in marine cultured fish.

The invention belongs to the technical field of poultry disease prevention and treatment, and discloses application of a rhizoma kaempferiae extract in preparation of a medicine for resisting avian infectious bronchitis virus. The rhizoma kaempferiae extract is prepared by taking rhizoma kaempferiae as a raw material and ethanol as an extraction solvent through extraction, filtration and concentration. Experimental results show that the rhizoma kaempferiae extract has no obvious toxicity to chick embryos within a certain dosage range; after the chicken embryo is infected with the infectious bronchitis virus, the survival rate of the chicken embryo can be improved by treating the chicken embryo with different doses of the rhizoma kaempferiae extract, and the rhizoma kaempferiae extract can reduce the virus antigen expression level and virus copy number in the liver tissue and allantoic fluid of the chicken embryo, so that the chicken embryo has prevention and treatment effects on infectious bronchitis virus infection. The rhizoma kaempferiae extract has the advantages of easily available raw materials, simple process and low cost, and is suitable for popularization and application.

This invention discloses an oral vaccine against carp spring viremia virus, its preparation method, and its application. The oral vaccine is constructed by linking a carp spring viremia virus antigen protein and a Yellow River carp defensin protein via a flexible linker to form a fusion protein, which is then expressed in *E. coli* and enriched using a biological vector. The gene sequence encoding this fusion protein includes a truncated fragment of the carp spring viremia virus antigen protein gene (64-1389 bp), a fragment of the Yellow River carp defensin protein gene (70-201 bp without the signal peptide), and a flexible linker fusion gene connecting the two gene fragments. Compared with previously reported anti-SVCV vaccines, the oral vaccine against carp spring viremia virus prepared in this invention does not harm the fish during the entire immunization process, is simple to operate, safe and stable, and has a higher protection rate. Therefore, this oral vaccine provides broad prospects for the prevention and control of carp spring viremia.

The invention provides a linear epitope of an African swine fever virus CP312R protein. The amino acid sequence of the linear epitope is SEQ ID NO: 1 or SEQ ID NO: 2. The antigen epitope polypeptide provided by the invention has good antigen specificity and immunoreactivity. The polypeptide can be efficiently combined with an African swine fever virus specific antibody, background interference is low, cross reaction is small, and detection sensitivity and accuracy are remarkably improved. By using the polypeptide as a coating antigen in immunodetection methods such as ELISA or colloidal gold immunochromatography, rapid and accurate detection of ASFV infection can be realized, and the detection result is stable and reliable. Compared with a traditional whole virus antigen or a crude extract antigen, the epitope polypeptide has the advantages of being simple and convenient to prepare, small in batch difference, high in safety and the like, and is suitable for ASFV serological monitoring and diagnosis application.

The invention belongs to the technical field of biology, and particularly relates to an evaluation method for the copy number of specific nucleic acid in a quantitative pseudovirus antigen standard substance, and the evaluation method comprises the following steps: (1) providing a pseudovirus antigen standard substance; (2) treating a pseudovirus antigen standard substance by DNA enzyme; (3) extracting nucleic acid of the treated pseudovirus antigen standard substance; (4) preparing a micro-droplet type digital PCR reaction system; (5) carrying out micro-droplet treatment on the reaction system; (6) performing micro-droplet digital PCR amplification on each reaction system subjected to micro-droplet treatment, and calculating the nucleic acid concentration of the pseudovirus antigen standard substance according to an amplification result; the invention also provides a primer probe composition for detecting rabies virus pseudovirus based on the disclosed method. According to the provided method and composition, accurate evaluation of the copy number of specific nucleic acid in a pseudovirus antigen standard substance is realized through the optimal primer and probe combination.

This application relates to the technical field of detection kits, and discloses an influenza virus antigen detection kit, which includes a box body, a cover plate, a handle, a detection platform, a placement groove opened at the top of the detection platform, and a detection strip placed in the placement groove. A driving device for driving the movement of the detection platform is installed in the box body. The driving device includes two first vertical grooves respectively opened on the inner walls of both sides of the box body, two first guide rods respectively fixed to the two first vertical grooves, and two sliding sleeves respectively slidably connected to the two first guide rods; the two sliding sleeves are respectively fixed to both sides of the detection platform. A first through hole is opened on one side of one first vertical groove, a first rack is slidably connected in the first through hole, and one of the sliding sleeves is fixed to one side of the first rack. A first gear is sleeved and fixed on the rotating shaft, and the first gear meshes with the first rack; a limiting mechanism for limiting the first rack is installed on the side wall of the first through hole. This application is convenient for testers to move and carry.

To provide a method for vaccinating a subject against mucosal viral infections.SOLUTION: The present disclosure provides a method for vaccinating a subject against a mucosal virus infection, comprising administering to the subject an immunologically effective amount of an intranasal booster, the intranasal booster comprising detoxified Escherichia coli labile toxin (LT) and an antigen from the mucosal virus, and the subject having been previously primed. An intranasal vaccine composition comprising an immunologically effective amount of a mucosal virus antigen adjuvanted with a detoxified Escherichia coli labile toxin (LT) is also provided.SELECTED DRAWING: Figure 1A

The invention provides a monoclonal antibody 3G1 capable of specifically recognizing CV-A5 virus and having neutralizing activity and application of the monoclonal antibody 3G1. The monoclonal antibody is prepared from a CV-A5 virus antigen immunized mouse and splenocytes of the mouse through a cell fusion technology, and amino acid sequences of three CDR regions of a heavy chain variable region of the monoclonal antibody sequentially comprise sequences as shown in SEQ ID NO.1-3; the amino acid sequences of the three CDR regions of the light chain variable region sequentially comprise sequences as shown in SEQ ID NO.4-6. The monoclonal antibody can be specifically combined with a CV-A5 virus, and is not combined with enteroviruses such as EV-A71, CV-A10, CV-A6, CV-A16 and the like. The monoclonal antibody targets conformational epitopes, has neutralizing activity, can specifically recognize CV-A5 viruses, is an ideal CV-A5 antigen detection antibody, and is beneficial to acceleration of the research and development process of hand-foot-mouth multivalent vaccines containing CV-A5 pathogens.

The present invention discloses a DC cell-based bifunctional nanobody, its construction method and application. The bifunctional nanobody is obtained by connecting a DC cell-based target protein-specific nanobody and a gene encoding a virus antigen-specific nanobody with a particle size of less than 150 nm through a linker element to obtain a target gene fragment and then recombinantly expressing the target protein. The technical means adopted in the present invention is universal, and specific nanobodies can be screened for different pathogens to construct corresponding porcine DC-targeted bifunctional nanobodies, which can be extended to other porcine vaccine antigens. The operation is simple, efficient, and time-consuming, and the bifunctional nanobody is relatively clear in design and structure, easy to be efficiently produced using a microbial genetic engineering system, with low manufacturing cost and good stability. The bifunctional nanobody can be used after incubation with the antigen, which is simple and easy to perform, more in line with the actual veterinary clinic, and convenient for popularization and application.

The invention belongs to the field of virus detection and diagnosis, and relates to an anti-AAV8 monoclonal antibody as well as a preparation method and application thereof. The invention provides an anti-AAV8 rabbit monoclonal antibody, and amino acid sequences of a heavy chain variable region and a light chain variable region of the anti-AAV8 rabbit monoclonal antibody. The anti-AAV8 rabbit monoclonal antibody provided by the invention can be specifically combined with VLP of AAV8, and can be used for detecting an AAV virus antigen. The anti-AAV8 monoclonal antibody provided by the invention provides an effective detection tool for AAV8 detection in a gene therapy vector research and development process and AAV8 detection in basic scientific research.

The present disclosure is directed to antibodies binding to and neutralizing henipavirus and methods for use thereof. Thus, in accordance with the present disclosure, there is provided a method of detecting a henipavirus infection in a subject comprising (a) contacting a sample from said subject with an antibody or antibody fragment having clone-paired heavy and light chain CDR sequences from Tables 3 and 4, respectively; and (b) detecting henipavirus in said sample by binding of said antibody or antibody fragment to a henipavirus antigen in said sample.