289 results about "Subunit vaccines" patented technology

The invention discloses a Graisseria parasuis three-component subunit vaccine and a preparation method thereof, and belongs to the technical field of biology. The vaccine comprises three kinds of antigen proteins of the Gleisseria parasuis in an immunizing dose and a pharmaceutically acceptable adjuvant, and the amino acid sequences of the three kinds of antigen proteins are respectively shown as SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3. The invention further discloses a preparation method of the vaccine. The three-component subunit vaccine provided by the invention has cross protection force on infection of type-4 and type-5 Graisseria parasuis, has an excellent immune protection effect, and is expected to play a better role in prevention and control of infectious diseases caused by the Graisseria parasuis.

The invention relates to the technical field of biology, in particular to a pseudorabies virus gD recombinant protein, a subunit vaccine and a preparation method and application thereof.The gD recombinant protein sequentially comprises a signal peptide, a connecting peptide, the 21st-350th site of the gD protein, an enzyme cutting site and a His tag from the N end to the C end, the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.4, and the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.6. The pseudorabies virus gD recombinant protein has high immunogenicity, the prepared subunit vaccine can generate a specific antibody with neutralizing activity, mice do not die after challenge, the protection rate is 100%, the protection rate after challenge of pigs is 100%, pathological sections only show slight pathological changes, and the pseudorabies virus gD recombinant protein has wide application prospects.

The invention belongs to the technical field of biology, and discloses application of a mycoplasma hyopneumoniae Ebh GA protein in preparation of mycoplasma hyopneumoniae or a multi-combined vaccine containing the mycoplasma hyopneumoniae. The applicant screens out the Ebh GA protein with immunogenicity from the mycoplasma hyopneumoniae for the first time, the Ebh GA protein is shown in SEQ ID NO.7 and can be used as a mycoplasma hyopneumoniae subunit vaccine, and therefore the applicant screens out the protein with immunogenicity of other pathogenic bacteria at the same time and combines the protein with the protein into a quadruple subunit vaccine. The antigens of the obtained quadruple vaccine have a synergistic effect, and compared with a single dose, the quadruple vaccine can enhance the immune effect of the quadruple vaccine.

The invention discloses a haemonchus contortus subunit vaccine, and belongs to the field of veterinary vaccines. The subunit vaccine comprises an antigen protein MEP3, an antigen protein AP1, an antigen protein AP8, an antigen protein H11-2, an antigen protein H11-4 and an antigen protein AP5 of galactosylated and modified haemonchus contortus, and the antigen protein MEP3, the antigen protein AP1, the antigen protein AP8, the antigen protein H11-2, the antigen protein H11-4 and the antigen protein AP5 of galactosylated and modified haemonchus contortus. The galactosylation modification of the antigen protein is realized by jointly expressing glycosyl transferase HcGALT of haemonchus contortus and one or more of the proteins in eukaryotic cells. The antigen protein modified by galactosylation in the subunit vaccine is closer to the glycosylation modification of natural protein, and can provide effective immune protection after the goat is infected with haemonchus contortus, and the egg laying rate is reduced by 81.43%. The invention lays a foundation for development of haemonchus contortus subunit vaccines, provides an effective technical means for prevention and treatment of the haemonchus contortus subunit vaccines, and has important application value and popularization prospect.

The present invention relates to the field of veterinary vaccines, and discloses the development and application of a subunit vaccine for porcine contagious pleuropneumonia. The TbpB-C truncated protein and the AdhE whole protein selected in the antigen protein combination of the vaccine of the present invention are highly homologous in all APP serotypes, and are prepared into subunit vaccines that can provide cross protection to porcine actinobacillus pleuropneumoniae of different serotypes, and the protection rates against APP1 type and APP3 type strains are both 100%. The truncated proteins selected in the antigen protein combination of the present invention are successfully expressed in a soluble form after optimization, and are easy to express and purify in large quantities, thereby reducing production costs. The present invention lays a foundation for the development of an efficient, low-cost, broad-spectrum subunit vaccine for porcine contagious pleuropneumonia, and provides an effective technical means for the prevention and treatment of the disease.

The invention belongs to the field of veterinary drugs, and relates to an avian infectious bronchitis virus antigen recombinant protein S-Trimer and a subunit vaccine thereof. The invention provides an avian infectious bronchitis virus antigen recombinant protein S-Trimer. The amino acid sequence of the antigen recombinant protein S-Trimer is as shown in SEQ ID No. 4. The invention provides an infectious bronchitis virus antigen. The infectious bronchitis virus antigen is of a trimer structure of the recombinant protein S-Trimer. The invention provides a chicken infectious bronchitis virus subunit vaccine. The subunit vaccine comprises a pharmaceutically acceptable carrier and an immune dose of the chicken infectious bronchitis virus antigen. The vaccine is high in safety, good in immunogenicity and stable in batches, and can provide complete protection for attacking the infectious bronchitis virus.

The invention provides a novel three-antigen HSV-2 adjuvant subunit vaccine as well as a preparation method and application thereof. The vaccine comprises herpes simplex virus recombinant protein and a composite adjuvant, the herpes simplex virus recombinant protein comprises extracellular domains of three proteins, namely HSV-2gE, gD and gC, wherein the amino acid sequences of the extracellular domains of the HSV-2gE, the gD and the gC are respectively shown as SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3; and the composite adjuvant is composed of CpG oligodeoxynucleotide and QS-21. Experiments prove that by adding the composite adjuvant composed of CpG oligodeoxynucleotide and QS-21, the immunogenicity, in-vivo protectiveness, virus removal and other immune functions of the HSV-2 subunit vaccine can be remarkably improved; meanwhile, the vaccine is a cross immune protection vaccine of an HSV-2 virus strain and an HSV-1 virus strain.

The invention discloses a swine fever and porcine parvovirus bivalent subunit vaccine and a preparation method thereof. The vaccine comprises a first recombinant protein encoded by a first gene, a second recombinant protein encoded by a second gene and a pharmaceutically acceptable carrier. The first gene has a sequence as shown in SEQ ID NO: 1 or an increased or reduced sequence thereof. And the second gene has a sequence as shown in SEQ ID NO: 2 or an increased or reduced sequence thereof. An antigen E2 protein of a hog cholera virus (CSFV) and a VP2 protein of a porcine parvovirus (PPV) are taken as double targets, a recombinant SC-E2 protein with a SpyCatcher tag and a recombinant ST-VP2 protein with a SpyTag tag are respectively expressed in insect cells through a recombinant baculovirus vector, double-antigen covalent assembly is realized in vitro, and the constructed bivalent subunit vaccine can be used for simultaneously preventing and controlling two epidemic diseases and has a good application prospect. And the vaccine has the advantages of high safety, strong immunogenicity, high prevention and control efficiency, easiness in large-scale production and the like.

The invention discloses a porcine delta-coronavirus double-antibody sandwich ELISA (enzyme-linked immuno sorbent assay) detection kit based on an S protein monoclonal antibody as well as a use method and application of the porcine delta-coronavirus double-antibody sandwich ELISA detection kit. A PDCoV S protein is expressed by adopting an ExpiCHO expression system, then the purified S protein is used as an immunogen to immunize a mouse, and the monoclonal antibody is obtained. And then, respectively using the monoclonal antibody 3D7 and the MAb 9G4 labeled by the HRP as capture and detection antibodies, and establishing the double-antibody sandwich ELISA kit. The kit has good sensitivity, the detection limit of purified S protein is 0.12 ng / mL, and the detection limit of PDCoV is 1.96 * 10 < 3 > copies / [mu] L. Specificity analysis shows that the kit has no cross reactivity with other porcine intestinal coronaviruses PEDV, TGEV and PoRV with clinical symptoms similar to PDCoV, the consistency with an RT-PCR (reverse transcription-polymerase chain reaction) method reaches 91.03%, and the Kappa value is 0.814, so that the reliability of the kit for clinical sample detection is shown. In addition, the kit can also quantitatively evaluate the content of S protein in PDCoV inactivated vaccines and subunit vaccines, and provides a powerful tool for evaluating the vaccines.

The invention relates to the technical field of aquaculture fish vaccines, in particular to a fusion gene of grass carp NPM1a and II type reovirus VP38, recombinant protein, a composition and application. The nucleotide sequence of the fusion gene is as shown in SEQ ID NO: 1. The fusion gene is obtained by connecting grass carp NPM1a and II type reovirus VP38 through a linker sequence, a BamH restriction enzyme cutting site is added to the 5'end of the fusion sequence, and an XhoI restriction enzyme cutting site is added to the 3 'end of the fusion sequence; the sequence of the linker is as shown in SEQ ID NO: 2. The subunit vaccine of the fusion protein of the grass carp NPM1a and the II type reovirus VP38 is prepared, and indole-3-lactic acid is used as an immunopotentiator in a combined manner, so that the technical bottlenecks of low protection rate, poor intestinal mucosal barrier repairability and the like of the existing vaccine are overcome.

The invention discloses construction and application of a streptococcus suis serotype 2 cps2C gene knockout mutant strain. The invention relates to a streptococcus suis serotype 2 cps2C gene knockout mutant strain 05ZYH33 delta cps2C. A coding gene, from the 31st site to the 1000th site, of the cps2C gene in the strain 05ZYH33 is replaced by a spectinomycin resistance gene cassette SpcR. The mutant strain 05ZYH33 delta cps2C is applied to preparation of a streptococcus suis type 2 attenuated vaccine and a subunit vaccine. The capsule of the mutant strain 05ZYH33 delta cps2C is obviously reduced; in-vitro experiments show that the macrophage phagocytosis resistance of the mutant strain 05ZYH33 delta cps2C is reduced, and animal toxicity test results show that the toxicity of the mutant strain is obviously reduced. The mutant strain provides an important clue for screening of protective antigens of multivalent subunit vaccines, and can be applied to development of S.suis attenuated vaccines and multivalent subunit vaccines.

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 invention provides a preparation method of a novel PEDV (porcine epidemic diarrhea virus) subunit vaccine, and belongs to the technical field of biology. The novel PEDV subunit vaccine is prepared by adopting a method comprising the following steps: (1) treating lactococcus lactis by using a high-pressure damp-heat method to obtain peptidoglycan particles; (2) treating the peptidoglycan particles by adopting an ultrahigh-pressure microjet technology to obtain a peptidoglycan carrier with the particle size of 280-850nm; and (3) mixing the peptidoglycan carrier with a PEDV subunit antigen of which the amino acid sequence is SEQ ID NO: 1 to obtain the novel PEDV subunit vaccine. The novel PEDV subunit vaccine comprises a peptidoglycan carrier with a specific size, can improve the in-vivo presentation efficiency of an antigen and rapidly induce efficient body fluid and mucous membrane immune response, and is of great significance to research and development of vaccines for porcine epidemic diarrhea diseases and the like.

The invention relates to a novel three-antigen HSV-2 subunit vaccine as well as a preparation method and application thereof, and belongs to the technical field of biology. The novel three-antigen HSV-2 subunit vaccine comprises antigens and a composite adjuvant, the antigens comprise HSV-2 gB2 envelope glycoprotein, HSV-2 gC2 envelope glycoprotein and HSV-2 gD2 envelope glycoprotein, and the composite adjuvant is CpG oligonucleotide and an aluminum adjuvant; the vaccine provided by the invention can induce a high-level neutralizing antibody, and widens targets for developing a novel multi-target antigen HSV-2 virus vaccine.

The present invention relates to a recombinant chimeric antigen targeting poxvirus (particularly monkeypox virus), a subunit vaccine thereof, and its application. The recombinant chimeric antigen of the present invention comprises two immunogens arranged in a specific manner: monkeypox virus A35 protein or its antigenic fragment (or its derivative peptide segment) and monkeypox virus M1 protein or its antigenic fragment (or its derivative peptide segment), which can stimulate immune responses against two infectious virus particles: intracellular mature virus particles (IMV) and extracellular enveloped virus particles (EEV), thereby efficiently stimulating specific immune protection against monkeypox virus. In addition, the poxvirus vaccine of the present invention also has good safety, rapid responsiveness and production capacity support, and has excellent clinical application prospects.

The invention discloses a chicken infectious anemia subunit vaccine composition as well as a preparation method and application thereof. The vaccine composition contains a mixture of CIAV (Chicken Infectious Anemia Virus) VP1 and VP2 proteins, the mixture of the VP1 and VP2 proteins is prepared by performing in-vitro renaturation on VP1 inclusion body proteins by using a gradient dialysis method, adding pre-purified VP2 proteins in the dialysis process to promote the recovery of key conformation epitopes of the VP1 proteins, and then gradually removing a denaturing agent to obtain the VP1 and VP2 protein mixture. And refolding the VP1 protein to obtain the protein. The soluble VP2 protein is used as a molecular chaperone to recover the VP1 protein from the inclusion body. According to the VP2 protein assisted synergistic refolding strategy, the key conformation epitope of the VP1 protein is recovered by using the natural function of the VP2 protein. Subsequently, the immunogenicity and protective efficacy of the refolded VP1 and VP2 protein mixture are evaluated, and an economic and effective technical means is provided for preventing and controlling chicken infectious anemia.

The invention discloses a paralichthys olivaceus rhabdovirus G protein tandem antigen epitope peptide and application thereof, and belongs to the field of fish molecular immunology. The amino acid sequence of the tandem antigen epitope peptide is shown as SEQ ID NO: 1. The preparation method comprises the following steps: (1) firstly, analyzing structural characteristics of HIRRV-G protein, predicting B cell antigen epitopes of the HIRRV-G protein, screening the antigen epitopes with advantages on the basis of a prediction result, and synthesizing the antigen epitopes; (2) screening a candidate peptide fragment with high affinity through an enzyme-linked immunosorbent assay; and (3) sequentially connecting the high-affinity peptide fragment sequences meeting the requirements by using a GPGPG connexon, cloning the connected sequences into a pET-28a prokaryotic expression vector, and performing induced expression to obtain the tandem antigen epitope peptide. Compared with a full-length G protein, the tandem antigen epitope peptide is smaller in molecular weight, higher in stability and higher in hydrophilicity; a large number of specific antibodies can be induced in fish bodies, and the death rate of the paralichthys olivaceus infected by viruses is remarkably reduced. The method can be used for HIRRV diagnosis detection reagent and subunit vaccine development.

The invention is applicable to the technical field of genetic engineering and biological medicine, and provides a construction method and an expression system of recombinant protein for preventing infectious bronchitis of chicken and application of the recombinant protein. Coding DNA of multi-antigen epitope tandem protein from infectious bronchitis virus N, S1 protein is connected to obtain a fusion recombinant protein IBV: NS1 coding gene, a recombinant expression plasmid pET28a-IBV: NS1 is constructed and transferred into E. coli Rosetta (DE3) competent cells to obtain an E. coli genetic engineering expression bacterium Ec-RD-pET28a-IBV: NS1, and the fusion recombinant protein IBV: NS1 is obtained through induced expression and massive purification. And mixing with an immunologic adjuvant to prepare the subunit vaccine. The subunit vaccine has the advantages of high antigen expression quantity, long antibody maintenance time, high purity, good safety, strong immunogenicity and strong protectiveness, and can effectively prevent infectious bronchitis.

This application discloses a feline allergen Fel d1 egg yolk antibody and its preparation method and application, comprising: obtaining PCR amplification products through PCR amplification; constructing a recombinant plasmid by ligating the PCR amplification products with pET-28a(+); transforming the recombinant plasmid into Rosetta competent cells to obtain transformants; obtaining recombinant antigens by extracting proteins from the transformants; obtaining a subunit vaccine by mixing and emulsifying the recombinant antigens with white oil adjuvant; administering the subunit vaccine to laying hens to obtain immunized egg yolk liquid; adding Tris-HCl buffer and polyethylene glycol to the egg yolk liquid and centrifuging to collect the protein aqueous phase; filtering the protein aqueous phase to obtain a filtrate, adding polyethylene glycol, mixing, centrifuging to collect the precipitate; mixing the precipitate with PBS buffer, adding ammonium sulfate, and centrifuging to obtain the egg yolk antibody. This invention solves the problem that the safety and economy of existing feline allergy treatment methods cannot be simultaneously achieved.

The invention discloses a recombinant 2.1 d subtype swine fever E2 protein, a subunit vaccine and application of the subunit vaccine, and belongs to the technical field of genetic engineering. According to the 2.1 d subtype swine fever E2 protein, 344-375 amino acids of the 2.1 d subtype swine fever E2 protein are removed, and 1-343 amino acids of the 2.1 d subtype swine fever E2 protein are reserved, so that the most important antigenic epitope with a protective effect of the 2.1 d subtype swine fever E2 protein can be reserved, the space structure of the recombinant 2.1 d subtype swine fever E2 protein is basically not influenced, and the immunogenicity of the 2.1 d subtype swine fever E2 protein can be reserved to the greatest extent; in addition, the recombinant 2.1 d subtype swine fever E2 protein can be efficiently expressed in a prokaryotic expression system; furthermore, the soluble expression level of the recombinant 2.1 d subtype swine fever E2 protein can be remarkably improved through co-expression of the molecular chaperone, and after the recombinant 2.1 d subtype swine fever E2 protein is prepared into a subunit vaccine, the subunit vaccine has a good protection effect on 2.1 d subtype swine fever virus infection.

The present application belongs to the field of veterinary drugs, and provides a recombinant protein S-Trimer of an infectious bronchitis virus antigen and a subunit vaccine therefor. Provided in the present application is a recombinant protein S-Trimer of an infectious bronchitis virus antigen, wherein the recombinant protein S-Trimer of the antigen has an amino acid sequence as set forth in SEQ ID No. 4. Also provided in the present application is an infectious bronchitis virus antigen. The infectious bronchitis virus antigen is of a trimer structure of the recombinant protein S-Trimer described above. The present application provides a subunit vaccine for an infectious bronchitis virus, wherein the subunit vaccine comprises a pharmaceutically acceptable carrier and an immunogenic amount of the infectious bronchitis virus antigen.

The invention belongs to the field of gene engineering, and provides a mycoplasma ovipneumoniae multi-epitope fusion protein and preparation and application thereof.The inventor firstly successfully predicts antigen epitopes of three virulence genes and performs tandem expression on the predicated antigen epitopes to prepare a fusion protein antigen, and the fusion protein antigen is used for preparing the mycoplasma ovipneumoniae multi-epitope fusion protein. A secondary structure, a tertiary structure, physicochemical properties (including hydrophilicity, stability, PI value and the like), antigenicity, sensitization and the like of the fusion protein are predicted through biological online software, and the success rate of a test is increased; meanwhile, the fusion protein connected in series with the three virulence genes of the mycoplasma ovipneumoniae is prepared into a subunit vaccine, and a good immune protection effect is also achieved. The recombinant mycoplasma ovipneumoniae multi-epitope fusion protein prokaryotic expression plasmid pET28a < + >-EHP is successfully constructed, tandem expression of mycoplasma ovipneumoniae on an escherichia coli prokaryotic system is achieved, and an expression product can obtain a good immune protection effect.

The invention is applicable to the technical field of genetic engineering and biological medicine, and provides construction of a fusion recombinant protein ABT for preventing brucellosis and application of the fusion recombinant protein ABT in preparation of protective vaccines. The fusion recombinant protein ABT is composed of an immunological enhancement antigen and a multi-epitope tandem antigen, wherein the immunological enhancement antigen (named as A) comprises a Brucella ribosome L7 / L12 protein and a PADRE sequence; the multi-epitope tandem antigen is formed by connecting a dominant B cell epitope part (named as B), a dominant Tc cell epitope part and a dominant Th cell epitope part (named as T) in series, wherein the dominant B cell epitope part (named as B) is derived from Brucella SurA, OMP31, BP26 and Trigger factor proteins. The ABT Brucella multi-epitope subunit vaccine prepared by mixing the fusion recombinant protein ABT and an immunologic adjuvant has the characteristics of good purity, high safety and strong immunogenicity, can stimulate an organism to generate a protective antibody, and has long antibody maintenance time.

The invention provides a subunit vaccine composition for porcine epidemic diarrhea, porcine delta coronal and porcine rotavirus as well as a preparation method and application of the subunit vaccine composition, and is characterized in that the subunit vaccine composition comprises prokaryotically expressed swine erysipelas filamentous bacillus SpaA protein, viral subunit protein and pharmaceutically acceptable adjuvants; wherein the amino acid sequence of the prokaryotically expressed swine erysipelas filamentous bacillus SpaA protein is as shown in SEQ No.1, and the viral subunit protein is selected from one or more of porcine epidemic diarrhea virus S protein, porcine delta coronavirus S protein and porcine rotavirus VP8 protein. The vaccine composition disclosed by the invention has the advantages of strong immunogenicity, good safety, no immune interference, high neutralizing antibody titer and long antibody duration.

The invention discloses a carrier protein with an adjuvant function and application thereof. The carrier protein with the adjuvant function has a sequence as shown in SEQ ID NO. 1. According to the carrier protein with the adjuvant function, a part of sequence is intercepted from the flagellin FliC of Burkholderia cepacia, so that the adjuvant activity of the flagellin is reserved, and the immunogenicity and antigenicity of the flagellin are reduced; when being applied to a fusion protein or a small-molecule drug, the fusion protein or the small-molecule drug can enhance the immunogenicity, so that the fusion protein or the small-molecule drug has important significance on subunit vaccines, polypeptide vaccines or small-molecule drug carriers.

This invention discloses an antigen and vaccine preparation based on dominant Brucella epitopes, and their application, belonging to the field of recombinant subunit vaccine technology. The antigen is composed of CTL epitopes, HTL epitopes, and B-cell epitopes tandemly, and its amino acid sequence is shown in SEQ ID NO.1. Based on screened dominant CTL epitopes, HTL epitopes, and B-cell epitopes of the Omp25 and Omp31 outer membrane proteins, this invention constructs a novel antigen fusion polypeptide. The vaccine prepared based on this polypeptide effectively activates a significant dual immune response, enhancing cellular immunity while inducing the body to produce high levels of specific antibodies. It also provides good protection against organ damage caused by bacterial infection, offering better immune response and protective efficacy against Brucella, laying the foundation for the development of Brucella recombinant subunit vaccines.

The present invention discloses a recombinant porcine circovirus type 3 trimer protein, a preparation method and an application thereof. The present invention uses bioinformatics methods and resources to predict the B cell antigen epitopes and T cell antigen epitopes of the PCV3Cap protein. Taking into account the stability and immunogenicity of the antigen epitopes, a recombinant porcine circovirus type 3 trimer protein based on the PCV3Cap protein antigen epitope is designed, and its efficient soluble expression in Escherichia coli is achieved. The expressed recombinant porcine circovirus type 3 trimer protein can be mass-produced and purified by Ni-NTA affinity chromatography. The purified recombinant porcine circovirus type 3 trimer protein can assemble into a stable trimer structure. The subunit vaccine prepared using the recombinant porcine circovirus type 3 trimer protein can induce experimental pigs to produce a high level of antibodies and has a significant protective effect on the experimental pigs. The recombinant porcine circovirus type 3 trimer protein designed by the present invention provides a new idea for the development of PCV3 vaccine.

The application discloses a tandem antigen epitope peptide of G protein of Paralichthys olivaceus rhabdovirus and application thereof, and belongs to the field of fish molecular immunology. The amino acid sequence of the tandem antigen epitope peptide is shown as SEQ ID NO:1. The preparation method of the application comprises the following steps: (1) first, analyzing the structural characteristics of HIRRV-G protein and predicting B cell antigen epitopes, and synthesizing the antigen epitopes with advantages based on the prediction results; (2) then, screening candidate peptide segments with high affinity through enzyme-linked immunosorbent assay; (3) sequentially connecting the high-affinity peptide segment sequences meeting the requirements by using a GPGPG linker, and cloning the connected sequences into a pET-28a prokaryotic expression vector, so that the tandem antigen epitope peptide is obtained after induced expression. Compared with the full-length G protein, the tandem antigen epitope peptide has smaller molecular weight, stronger stability and stronger hydrophilicity; the tandem antigen epitope peptide can induce a large amount of specific antibodies in fish bodies, and significantly reduces the mortality of fish infected by the virus. The tandem antigen epitope peptide can be used for the development of HIRRV diagnostic reagents and subunit vaccines.

The present disclosure belongs to the field of biotechnology, and specifically relates to a seven-component antigen African swine fever subunit vaccine. The present disclosure first provides an African swine fever virus antigen protein combination composed of the African swine fever virus P34 protein, P30 protein, P54 protein, A104R protein, C129R protein, X protein, and Y protein. This African swine fever virus antigen protein combination can induce a strong immune response in the host. Furthermore, the present disclosure provides a seven-component antigen African swine fever subunit vaccine including the aforementioned African swine fever virus antigen protein combination. The seven-component antigen African swine fever subunit vaccine exhibits good immunoprotection rates against challenge with the parental virulent African swine fever virus strain, poses no biosafety risks, overcomes the difficulty that existing African swine fever subunit vaccines domestically and internationally cannot provide effective immunoprotection for pigs.

The invention discloses a bivalent subunit vaccine for preventing infection of haemophilus parasuis and porcine circovirus type 2. A core antigen combination of the vaccine comprises at least one antigen protein from haemophilus parasuis, and the antigen protein is selected from Ferrin, OppA and Hem-SAP and is combined with a porcine circovirus type 2 Cap protein. Wherein the NCBI (National Center of Biotechnology Information) login number of the Ferrin, the NCBI login number of the OppA and the NCBI login number of the Hem-SAP are WP160414389.1, ACL32731.1 and WP035493594.1 respectively. The antigen protein is subjected to codon optimization, is expressed and purified through a prokaryotic expression system, and is emulsified with ISA201 or a Freund's adjuvant to prepare the vaccine. Animal experiments prove that the vaccine can excite high-level antigen specificity IgG, cell factors IFN-gamma and white IL-4 in an immune animal body, namely, specific Th1 and Th2 type immune responses are generated. The vaccine can generate an immune protection rate of up to 80% when attacked by a serum type 5 haemophilus parasuis virulent strain, and generates an effective antibody response to the porcine circovirus type 2. The vaccine provided by the invention has the advantages of definite components, good safety and strong immune protection force.