183results about How to "Improving immunogenicity" patented technology

The invention provides a porcine epidemic diarrhea virus S protein mutant, a nucleic acid molecule, a subunit vaccine and application. Compared with wild type porcine epidemic diarrhea virus S protein, the porcine epidemic diarrhea virus S protein mutant has the advantage that amino acid at the 1076th site, amino acid at the 1077th site, amino acid at the 1016th site and amino acid at the 1017th site are mutated into proline. The natural trimer which is stable and maintains the conformation of the S protein before fusion is constructed, the safe and efficient PEDV subunit vaccine is obtained, and the use effect and immunogenicity of the porcine epidemic diarrhea vaccine can be improved; moreover, compared with a wild type S protein, the porcine epidemic diarrhea virus S protein mutant disclosed by the invention has a higher expression quantity.

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.

The invention discloses a chimeric PEDV (porcine epidemic diarrhea virus) infectious clone plasmid, a PEDV chimeric virus as well as a construction method and application thereof. According to the invention, a PEDV genome is used as a skeleton for the first time, the chimeric virus for expressing the extracellular domain of the TGEV S protein is successfully rescued, and it is found that the TCID50 of the chimeric virus for expressing the extracellular domain of the TGEV S protein can be increased by 6.2 times compared with that of the chimeric PEDV for expressing the TGEV S protein. The recombinant virus is used as an inactivated vaccine for immunizing mice, so that the immunogenicity of TGEV S protein can be greatly improved, a body is stimulated to generate a higher-level neutralizing antibody aiming at TGEV, and the titer of the neutralizing antibody aiming at TGEV is improved by 16.8 times. Therefore, more possibilities are provided for modification of viral genomes, a novel method for improving immunogenicity of the TGEV inactivated vaccine is invented, and a basis is provided for research and development of the TGEV vaccine.

The invention discloses a mycoplasma hyopneumoniae multi-antigen fusion protein and a vaccine composition and application thereof, and belongs to the technical field of genetic engineering and vaccines. The fusion protein comprises an escherichia coli heat-labile enterotoxin LTB, a mycoplasma hyopneumoniae antigen protein P97R1, a mycoplasma hyopneumoniae lipoprotein 418 and a mycoplasma hyopneumoniae surface antigen protein P46 from the N terminal to the C terminal in the sequence. The vaccine composition comprises the fusion protein and an adjuvant acceptable in veterinary medicine. A vaccine prepared from the fusion protein has the advantages of higher pertinence and immunogenicity, the cell and humoral immune response level is remarkably improved, and meanwhile, the fusion protein is easy to purify and prepare and suitable for large-scale industrial production.

The invention discloses a preparation method and application of porcine pseudorabies virus multi-epitope polypeptide, and belongs to the technical field of veterinary biological products. The multi-epitope polypeptide comprises a cytotoxic T cell epitope, a helper T cell epitope and a linear B cell epitope; the cell epitopes are screened from key immunogenic proteins of the pseudorabies virus by using an immunoinformatics method. The porcine pseudorabies virus multi-epitope polypeptide designed by the invention is high in safety, has better antigenicity and physicochemical properties, can be efficiently expressed, and can be recognized by a pseudorabies virus specific antibody. By applying the porcine pseudorabies virus multi-epitope polypeptide, high-level specific antibodies and cellular immune response can be continuously induced to generate, strong protective efficacy is generated, and mice can be protected from death when infected with a lethal dose of porcine pseudorabies virus. Therefore, the multi-epitope polypeptide disclosed by the invention can be used as an alternative antigen of a porcine pseudorabies virus vaccine, the use of active virus is avoided, and a tool is provided for eradication of porcine pseudorabies virus.

The invention relates to the technical field of bioengineering, and particularly discloses a chicken infectious anemia virus, a chicken infectious anemia virus egg yolk antibody and a preparation method and application thereof. The invention relates to a chicken infectious anemia virus, which is preserved in the China General Microbiological Culture Collection Center (CGMCC), and is named as the chicken infectious anemia virus RD240101A, the preservation date is September 23, 2024, and the preservation number is CGMCC NO.46198. The chicken infectious anemia virus has the advantages that the chicken infectious anemia virus is named as the chicken infectious anemia virus RD240101A; the chicken infectious anemia virus egg yolk antibody is prepared by taking an inactivated vaccine prepared from the chicken infectious anemia virus RD240101A as an immunogen, is high in specificity and good in safety, does not have any local or whole-body adverse reaction caused by injection of the antibody after being inoculated to chicks, can provide effective immune protection for chicken flocks, and can be used for preparing the chicken infectious anemia virus egg yolk antibody. And the method has a good commercial development prospect.

The invention provides a chicken infectious laryngotracheitis subunit vaccine and application thereof, and belongs to the technical field of vaccine preparation. The chicken infectious laryngotracheitis subunit vaccine provided by the invention is prepared from chicken infectious laryngotracheitis virus gB-A463P protein coded by a gB-A463P gene of which the 463th alanine of an optimized chicken infectious laryngotracheitis gB gene is further mutated into proline, and the amino acid sequence of the gB-A463P protein is shown as SEQ ID NO: 5. The subunit vaccine is prepared by adopting a genetic engineering means, only contains part of antigens of the ILTV and does not contain genetic materials, so that the risks of toxin expelling and toxin dispersing do not exist, and the safety can be effectively improved.

The invention relates to a preparation method of a tumor microenvironment response type nano drug delivery system, which comprises the following steps: forming MnO2 nanoclusters through biomineralization induction under an alkaline condition, modifying HSA nanoparticles, then loading Ce6 through a carbodiimide method, and finally combining with a PD-L1 antibody by adopting a copper-free click chemistry method to obtain nanoparticles HMC and anti-PD-L1. The nanoparticle can be retained in a tumor tissue in a passive targeting manner through an EPR effect, and is decomposed into Mn < 2 + >, O2 and a small molecule drug through a high H2O2 and low pH microenvironment of the tumor tissue, so that the tumor permeability is greatly enhanced, the tumor immunogenicity is enhanced by combining sonodynamic therapy, the release of a tumor specific antigen and the aging of antigen presenting cells are promoted, and the tumor treatment effect is enhanced. The infiltration of tumor T cells is increased, and cold tumors are changed into hot tumors; the nanoparticles can generate O2 in situ at the same time, and the immunosuppressive ability and drug resistance caused by tumor hypoxia are overcome.

The invention relates to the technical field of tissue engineering, and particularly discloses preparation of an exosome and electrostatic spinning composite material for promoting bone regeneration and application of the exosome and electrostatic spinning composite material in bone tissue regeneration. Identifying the multiplication capacity, the surface marker and the multidirectional differentiation capacity of the cells after the cells are separated and cultured, and finally extracting and identifying the exosome; pLLA and gelatin are dissolved in a cosolvent trifluoroethanol, composite fibers are prepared through electrostatic spinning, and a composite fiber membrane is obtained through spinning by adopting a receiver; the exosome is loaded on the surface of the PLLA / gelatin composite fiber membrane. According to the invention, the exosome derived from human mesenchymal stem cells is combined with the membrane material to prepare the exosome capable of stably releasing, the good stability and low immunogenicity of the exosome are exerted, and the effect of accurately promoting local bone regeneration is realized. And defect repair and functional reconstruction are realized.

The invention discloses an antibody targeting a nerve cellosilk light chain as well as a preparation method and application of the antibody. According to the present invention, with the HEAD structure, the prokaryotic recombinant nerve fiber filament light chain antigen has strong immunogenicity, more dominant epitopes are exposed, and the monoclonal antibody with high affinity is prepared through immunization; besides, the HEAD structural domain is fused with the specific label on the carrier, so that the stability of the antigen is greatly improved, the problem of polysinolysis caused by the property of the antigen is solved, and the success rate of antibody preparation is further improved. The antibody or the antigen binding fragment thereof can be used for early diagnosis and prognosis of neurodegenerative diseases.

The invention belongs to the field of veterinary biological products, and particularly relates to a chicken infectious laryngotracheitis recombinant subunit vaccine as well as a preparation method and application thereof. A core antigen of the vaccine is a recombinant gB trimer glycoprotein (gB-Trimer) modified by genetic engineering, and the protein successfully locks a prefusion conformation with the strongest immunogenicity of the gB protein through strategies of truncation, trimerization motif splicing, dual-stability mutation and the like. The vaccine prepared by the invention can stimulate rapid and lasting neutralizing antibody response, provide 100% clinical protection and effectively block replication and detoxification of the ILTV, has extremely high biological safety compared with a traditional live vaccine, completely eliminates risks of poison dispersion and enhancement, and provides a safe and efficient novel solution for prevention and control of the ILTV.

The present invention relates to a vaccine framework fusion protein, a recombinant vaccine and its application. The fusion protein of the present invention comprises the intracellular segment of the low-affinity immunoglobulin gamma Fc region receptor II (FcγRII) and the WW domain (WW domain) of the E3 ubiquitin protein ligase Nedd4 family member protein, and / or one or more antigens, and / or one or more transmembrane regions. The present invention provides a new recombinant vaccine molecular architecture, which has a better ability to promote the expression and secretion of immune antigens and improve the effect of immune level, provides new ideas and methods for the design of recombinant vaccines, and can be widely used in the field of vaccines.

The invention discloses a foot and mouth disease virus effective antigen inactivation protective agent and application thereof. The protective agent is prepared from the following components in percentage by mass: 0.05 to 20 percent of a structure curing agent, 0.01 to 5 percent of an ion regulator, 0.005 to 1.0 percent of an antioxidant and the balance of water, the protective agent does not influence the inactivation effect in the process of acting on nucleic acid, inhibits folding and unfolding of a protein physical structure, prevents change of a protein space physical structure, improves free energy of protein denaturation, enhances the stability of a virus space physical structure, maintains the integrity of virions, and has a good application prospect. The effects of preserving the immune biological activity of the protein and reducing the loss of viruses in the inactivation process are achieved, so that the foot-and-mouth disease inactivation antigen liquid with high immunogenicity is obtained. Compared with the conventional antigen protection method, the method has the advantages that the virus is protected before the inactivation process is implemented, the production process is advanced, the stability of the effective antigen can be protected to the greatest extent, and support is provided for obtaining an effective antigen solution with high immunocompetence.

The invention relates to the technical field of biology, and particularly discloses a recombinant vector, a recombinant baculovirus and a preparation method and application of the recombinant vector and the recombinant baculovirus. According to the method for preparing the recombinant vector, pFastBac Dual is adopted as a vector to prepare a recombinant plasmid vector inserted into a VP2 gene of an avian bursal disease virus classic strain, and the recombinant plasmid vector is named as pDual-V1; the method comprises the following steps: preparing a recombinant plasmid vector into which VP2 genes of a classical strain and a variant strain of the avian bursal disease virus are inserted at the same time, and naming the recombinant plasmid vector as pDual-V1-V2; the method comprises the following steps: preparing a recombinant plasmid vector inserted into a group I type 4 fowl adenovirus Fiber-2 gene, and naming the recombinant plasmid vector as pDual-F2; the method comprises the following steps: preparing a recombinant plasmid vector into which a Fiber-2 gene and an avian reovirus sigma C gene sequence are inserted at the same time, wherein the recombinant plasmid vector is named as pDual-F2-sigma C; carrying out double enzyme digestion on the pDual-V1-V2, recovering small fragments, carrying out double enzyme digestion on the pDual-F2-sigma C, recovering large fragments, and then carrying out mixed enzyme ligation. The recombinant baculovirus can express four antigens at the same time, and effective preparation of a multi-combined vaccine is achieved.

The invention discloses a composite adjuvant, and belongs to the field of vaccine adjuvants. The main active components of the composite adjuvant are squalene, polysorbate 80 and monophosphoryl ester A, the composite adjuvant is in an oil-in-water emulsion form, nano-scale emulsion droplet particles are uniformly distributed in a water phase, and the system is stable. The invention also discloses a preparation method of the composite adjuvant based on a stirring method. The composite adjuvant comprehensively utilizes the dual characteristics of the oil-in-water emulsion and the immunopotentiator, and can induce a higher level of immunoreaction when being combined with the antigen, so that the composite adjuvant can be used for preparing vaccines and improving the effectiveness of the vaccines.

The invention provides a respiratory syncytial virus (RSV) F protein comprising at least one modification or mutation selected from the group consisting of: (1) introducing one or more amino acid mutations into the RSV F protein for filling a hydrophobic cavity; (2) introducing one or more amino acid mutations for electrostatic modification into the RSV F protein; and (3) removing a p27 peptide fragment in the RSV F protein. As a recombinant protein antigen, the RSV F protein is high in stability and easy to store and transport, the expression quantity is increased, the yield is easy to increase, and the modified RSV F protein is higher in immunogenicity and can generate a better immune protection effect.

The invention particularly relates to a diphtheria toxin antigen modified by fatty acid and application thereof, and belongs to the field of biological medicine, in the diphtheria toxin antigen modified by fatty acid, the fatty acid modification comprises N-terminal cysteine (Cys) modified by fatty acid, the N-terminal Cys modified by fatty acid is characterized in that sulfydryl containing Cys is modified by diacylglycerol and / or alpha amino is modified by fatty acyl. Compared with the existing diphtheria toxin protein antigen, the fatty acid modified diphtheria toxin antigen not only has faster induction and higher immune protection effect than the conventional intramuscular injection induction of the existing diphtheria toxin antigen, but also can induce respiratory tract mucosa immunity, and provides a basis for blocking diphtheria infection and diffusion propagation and providing a vaccine for immunoprophylaxis.

The invention discloses a porcine rotavirus RNA vaccine and a preparation method thereof. According to the invention, amino acid sequences and nucleotide sequences of G9 type and G4 type VP4 proteins of porcine rotaviruses are optimized, and optimized translation starting elements are used, so that optimized RNA molecules with nucleotide sequences shown as SEQ ID NO.18 and SEQ ID NO.24 are finally prepared. The optimized RNA has higher immunogenicity in a mouse body. Furthermore, the porcine rotavirus RNA vaccine provided by the invention comprises a vaccine vector and the optimized RNA molecule. The porcine rotavirus G9-G4 type bivalent RNA vaccine disclosed by the invention is superior to a porcine rotavirus inactivated vaccine in the aspect of immune protection efficacy in a pig body.

The invention relates to a mycobacterium tuberculosis vaccine, in particular, the vaccine comprises an immune composition, and the immune composition comprises an antigen component and a granular protein component. The granular protein component comprises nano-granular protein, and the antigen component and the granular protein component are covalently bound through a binding peptide 1 and a binding peptide 2 to form an immunogenic complex. The cell immunogenicity and the antibody immunogenicity of the vaccine are excellent. The invention also discloses a preparation method of the mycobacterium tuberculosis vaccine.

The invention belongs to the technical field of biological medicines, and particularly relates to a PPV-PCV2 nanoparticle vaccine design and a preparation method. According to the invention, PPV1VP2 protein is used as a skeleton, a SpyTag peptide fragment is inserted into a loop 2 region of the VP2 protein, and the fusion protein is expressed by using an insect cell baculovirus system and is marked as PPV-ST; a plurality of B cell epitopes of Cap protein of PCV2 and a plurality of T cell antigen epitopes of Rep protein of PCV2 are connected in series, a SpyCatcher peptide fragment is fused at an N end, and expression is performed by using a prokaryotic expression system and is marked as SC-PCV2. Finally, the PPV-ST protein and the SC-PCV2 protein are subjected to covalent binding assembly, the nanoparticle vaccine is prepared, and the obtained PPV-PCV2 vaccine has good immunogenicity.

The invention relates to a multi-antigen monkey pox RNA vaccine and a preparation method thereof, and relates to the technical field of nucleic acid vaccines. The multi-antigen mRNA comprises a sequence encoding an antigenic polypeptide of the monkey pox virus or an antigenic fragment, variant or derivative thereof, the antigenic polypeptide or the antigenic fragment thereof comprising at least viral proteins of an intracellular mature virus and an extracellular envelope virus of the monkey pox virus. The invention provides a multi-antigen mRNA vaccine aiming at monkey pox viruses and a sequence, mRNA can express antigen protein in in-vitro cells, high-titer anti-monkey pox virus A29L, A33L, B6R, M1R and E8L protein IgG can be detected after a mouse is immunized, a high-titer anti-vaccinia Tian Tan strain live virus neutralizing antibody is generated, good immunogenicity is achieved, and the vaccine can be used for preparing the multi-antigen mRNA vaccine aiming at the monkey pox viruses. The important significance is realized on the prevention of the monkey pox virus.

The invention belongs to the field of biological medicine, and particularly relates to application of LMP2 antigen mRNA. According to the LMP2 antigen mRNA design, an MHC-I molecular intracellular sequence is adopted, and the efficiency that an expression antigen is presented by MHC-I and MHC-II can be remarkably improved. The method has universality, the designed LMP2 antigen enhanced mRNA has high immunogenicity, and the prepared mRNA vaccine can be used for treating a series of cancers and tumors which are represented by nasopharynx cancer and are caused by virus infection.

The application discloses a fusion protein and a preparation method and application thereof. The fusion protein comprises a P1 protein C-terminal fragment and an adjuvant protein, and can be used for preparing a Mycoplasma pneumoniae recombinant protein vaccine. The P1 antigen fragment is fused and expressed with a mucosal adjuvant, so that high expression quantity, strong immunogenicity and induction of respiratory mucosal immune response are realized.

Disclosed are a recombinant fusion protein derived from an HR region of an S2 protein of SARS-CoV-2 and an application of the recombinant fusion protein. The SARS-CoV-2 recombinant fusion protein is a recombinant fusion protein obtained by linking two membrane fusion-related conserved amino acid sequences HR1 and HR2 of the SARS-CoV-2 membrane protein S2 protein by means of a linking peptide. The recombinant fusion protein can be induced and expressed in Escherichia coli, has high expression quantity, and is easy to purify. The SARS-CoV-2 recombinant fusion protein provided by the present invention can form and maintain a stable dimer structure, simulates the conformation of a SARS-CoV-2 membrane fusion intermediate state, can be used as a detection raw material for detecting a SARS-CoV-2 membrane fusion process, has good anti-SARS-CoV-2 activity and good immunogenicity, and has a wide application prospect in the fields of development of drugs for preventing or treating SARS-CoV-2 proteins and development of SARS-CoV-2 vaccines and anti-SARS-CoV-2 antibodies.

The present invention discloses a G9P[23] genotype porcine rotavirus strain and its application; the G9P[23] genotype porcine rotavirus is isolated and passaged from a piglet diarrhea sample in a pig farm in Guangdong Province, named RVA / pig / CH / U945 / 2022 / G9P[23], and the deposit number is CCTCC NO: V202454. The isolated strain can stably proliferate in MA104 cells and produce typical cytopathic effects. The G9P[23] genotype porcine rotavirus strain isolated by the present invention can be used for the preparation of virus strains for testing and inactivated vaccines. As a vaccine strain, it has good immunogenicity and can produce a high level of antibodies after immunizing animals, providing the necessary material and theoretical basis for the subsequent research and development of rotavirus vaccines.

The invention relates to the technical field of biological pharmacy, in particular to a multivalent HPV mRNA vaccine and a preparation method thereof. The present invention provides self-replicating and / or non-self-replicating mRNA molecules. The mRNA molecule provided by the invention is used for connecting E6 and E7 genes from HPV16 and HPV18, and four antigens can be expressed at the same time. After a therapeutic mRNA molecule obtained by combining the polypeptide with other elements of an mRNA vaccine is encapsulated by lipid nanoparticles, E6 and E7 four antigens of HPV16 and HPV18 can be simultaneously and successfully expressed in vivo and in vitro. When the vaccine is used for immunizing mice, higher cellular immune level and tumor curative effect can be generated, and the vaccine has a wide application prospect.

The invention discloses a hapten for detecting the content of isofenphos-methyl and application of the hapten, particularly relates to the hapten for detecting the content of isofenphos-methyl and a preparation method of the hapten, and further discloses application of the hapten in the aspects of antibodies and kits. The enzyme linked immunosorbent assay kit established by aiming at the hapten for detecting the content of isofenphos-methyl can realize rapid detection of a large batch of samples to be detected, is convenient to use and low in detection cost, the detection method is efficient, accurate and rapid, can simultaneously detect a large batch of samples, and is suitable for on-site monitoring of isofenphos-methyl in fruits, vegetables and traditional Chinese medicinal materials and screening of a large number of samples.

This invention relates to the field of biomedical technology, specifically to HCBP6 monoclonal antibodies and their applications. This invention utilizes bioinformatics databases and online tools to analyze the structure of the HCBP6 protein, screening for highly immunogenic HCBP6 antigen peptides. Antibody 1 and Antibody 2 are obtained through immunization. The combination of Antibody 1 and Antibody 2 is used for the detection of HCBP6, exhibiting high sensitivity and good detection results. It has significant development and application value and long-term implications for the auxiliary diagnosis of fatty liver.

The invention discloses an immunogenic Mycoplasma ovipneumoniae HSP70-P113 fusion protein, which is composed of a Mycoplasma ovipneumoniae heat shock protein gene HSP70 and Mycoplasma ovipneumoniae adhesin gene P113 The spliced ​​fusion gene encodes an amino acid sequence shown in SQE ID NO. 2. Western Blot test proves that the Mycoplasma ovipneumoniae HSP70-P113 fusion protein has immunogenicity and can be used to prepare Mycoplasma ovipneumoniae subunit vaccine.

The invention discloses a preparation method and application of a hexavalent multi-epitope fusion protein of avian pathogenic Escherichia coli. The invention preferably selects antigenic epitopes of the curli fimbriae subunit CsgA, lipoprotein SlyB, maltose porin LamB, outer membrane protein PhoE, proteolytic enzyme activator NlpD and lipoprotein Pal main structural subunit of avian pathogenic Escherichia coli, and preferably connects with the heat-labile enterotoxin LTb fusion protein in series to construct an antigenic multi-epitope fusion protein of six proteins of avian pathogenic Escherichia coli. The antigenic multi-epitope fusion protein has a wide serotype coverage, is non-allergenic, has strong immunogenicity, and produces strong specific humoral immunity and cellular immune response after immunizing animals.