33 results about "E2 protein" patented technology

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 relates to a monoclonal antibody DIDA55 aiming at hog cholera virus, and a preparation method and application thereof, belonging to the field of medical preparations. The monoclonal antibody DIDA55 or an antigen binding fragment thereof comprises a heavy chain variable region and a light chain variable region, amino acid sequences of LCDR1, LCDR2 and LCDR3 in a light chain variable region of the antibody are shown as 24th to 34th, 50th to 56th and 89th to 97th in SEQ ID No: 4; amino acid sequences of HCDR1, HCDR2 and HCDR3 in a heavy chain variable region of the antibody are shown as the 31st to 35th, the 50th to 65th and the 95th to 102th of SEQ ID No: 5. The monoclonal antibody provided by the invention can be used in the fields of swine fever E2 protein labeled subunit vaccine immunity, serological diagnosis of wild strains, swine fever vaccine immune effect evaluation, related experiments and the like, and provides antibody resources for prevention, control and purification of CSF.

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 invention provides a kit for detecting bovine viral diarrhea virus and application of the kit. According to the present invention, the BVDV E2 protein is optimized, the monoclonal antibody 3G5 capable of specifically identifying DVDV1 type and DVD2 type viruses is screened on the basis, and the monoclonal antibody not only can specifically identify the purified and expressed BVDV E2 protein, but also can specifically identify the BVDV in the natural virus infected MDBK cell. By preparing the monoclonal antibody of the BVDV-E2 protein, a series of BVDV immunological diagnostic reagents can be researched and developed, and the monoclonal antibody has important significance on BVDV diagnosis.

The invention discloses a chikungunya virus envelope E2 protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

The invention provides a bivalent bovine viral diarrhea virus antigen protein composition, a subunit vaccine as well as a preparation method and application of the bivalent bovine viral diarrhea virus antigen protein composition and the subunit vaccine, and belongs to the technical field of genetic engineering vaccines. The bivalent bovine viral diarrhea virus antigen protein composition is prepared from a recombinant protein kozak-MLTsp-BVDV1-E2-His and a recombinant protein kozak-IgKsp-BVDV2-E2-His, the amino acid sequence of the recombinant protein kozak-MLTsp-BVDV1-E2-His is as shown in SEQ ID NO.16, and the amino acid sequence of the recombinant protein kozak-IgKsp-BVDV2-E2-His is as shown in SEQ ID NO.18. The invention further discloses a preparation method of the bivalent bovine viral diarrhea virus antigen protein composition. According to the invention, efficient expression of BVDV-1 and BVDV-2 E2 proteins is realized, high-purity E2 proteins are obtained, the safety and effectiveness of vaccines are ensured, and the antibody level of the vaccines is improved.

The invention belongs to the technical field of biology, and particularly relates to a bispecific antibody targeting African swine fever virus and hog cholera virus and application of the bispecific antibody, the bispecific antibody shows high binding capacity to p30 protein of the African swine fever virus and E2 protein of the hog cholera virus, and it is indicated that the bispecific antibody can recognize the African swine fever virus and the hog cholera virus at the same time. When the bispecific antibody disclosed by the invention is used for preparing a detection product, double-flux detection of screening African swine fever viruses and swine fever viruses can be realized at the same time through one-time detection, and a new thought is provided for multiple detection of swine disease serology.

The embodiment of the present application relates to a codon-optimized bovine viral diarrhea virus type 1 E2 protein gene and application thereof, and belongs to the field of bioengineering, wherein the nucleotide sequence of the codon-optimized bovine viral diarrhea virus type 1 E2 protein gene is shown as SEQ ID NO:1. A CHO cell strain stably expressing bovine viral diarrhea virus E2 protein is successfully constructed, and the E2 protein expressed by the cell strain has the characteristics of high expression amount, good immunogenicity, easy purification and production.

The present application relates to a kind of monoclonal antibody DIVA55 for swine fever virus in the field of medical preparation and its preparation method and application.The monoclonal antibody DIVA55 or its antigen-binding fragment of the present application includes heavy chain variable region and light chain variable region;The amino acid sequences of LCDR1, LCDR2 and LCDR3 in the light chain variable region of the antibody are as shown in the 24th to 34th, 50th to 56th, 89th to 97th of SEQ ID No:4;The amino acid sequences of HCDR1, HCDR2 and HCDR3 in the heavy chain variable region of the antibody are as shown in the 31st to 35th, 50th to 65th, 95th to 102nd of SEQ ID No:5.The monoclonal antibody of the present application can be used in the field such as swine fever E2 protein marker subunit vaccine immunization and wild strain serological diagnosis, swine fever vaccine immunization effect evaluation and related experiments, provides antibody resource for CSF prevention and control and purification.

The application discloses an E2 fusion protein, a coding gene thereof and application of the E2 fusion protein. The E2 fusion protein is fused by an E2 protein and a label; the amino acid sequence of the E2 protein is shown as positions 1 to 365 from the N-terminal end in SEQ ID NO:1. Experiments prove that the E2 protein has a helicase activity, and an anti-human papillomavirus drug, a podophyllotoxin, can inhibit the helicase activity of the E2 protein. Therefore, the E2 protein is a target of the action of the podophyllotoxin, and the E2 protein can also be used as a target for screening the anti-human papillomavirus drug. The application has important application value in the process of screening the anti-human papillomavirus drug.

The application discloses a carbon-coated zirconium oxide composite material, a colorimetric / photothermal dual-mode signal nano probe and application thereof in rapid quantitative detection of chikungunya virus. The colorimetric / photothermal dual-mode signal nano probe is formed by coupling a carbon-coated zirconium oxide composite nano material obtained by calcining mesoporous UiO-66 in an inert atmosphere with a chikungunya virus E2 protein monoclonal antibody. In a specific application, a chromatography test strip is constructed based on the colorimetric / photothermal dual-mode signal nano probe. The chromatography test strip comprises a nitrocellulose membrane provided with a T line and a C line. During detection, a sample and the nano probe form an immune complex and generate a colorimetric signal visible to the naked eye at the T line, and meanwhile, under the irradiation of 808 nm near-infrared laser, a temperature rise Delta T related to the concentration of a target object is generated at the T line, so that on-site rapid screening and photothermal quantitative review are realized. The chromatography test strip has both simple operation and reliable quantification, and has stronger anti-interference ability in a complex biological matrix, and is suitable for instant detection of chikungunya virus.

The present invention relates to the field of vaccination, in particular, of vaccination against hepatitis C virus (HCV). The present invention provides a composition comprising at least parts of the HCV core protein, and HCV E1 and HCV E2 protein of a specific HCV strain, as well as VSV-G protein. The proteins may be assembled in rVSV-HCV particles. This has been identified to induce particularly advantageous broadly neutralizing antibodies. The invention further provides nucleic acids encoding said HCV proteins and VSV proteins but not encoding VSV-G protein. Vaccines comprising the particles, compositions or nucleic acids are disclosed as useful, in particular, for prophylactic vaccination against HCV. Methods of producing the rVSV-HCV particles or compositions of the invention and the produced particles and compositions are also subject-matter of the invention.

The invention discloses a chikungunya virus envelope E2 protein monoclonal antibody and application of the chikungunya virus envelope E2 protein monoclonal antibody in preparation of preventive and therapeutic antibody drugs, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

The present disclosure provides for novel nucleic acid constructs and methods for expressing HCV E2 protein on a cell surface as an active antigen. The present disclosure provides for a method for evaluation of protein expression. Vaccine formulations, constructs and expression vectors are all within the scope of this disclosure.

The invention provides a subunit vaccine composition for swine fever, porcine pseudorabies, porcine circovirus and swine erysipelas as well as a preparation method and application of the subunit vaccine composition. 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 subunit E2 protein of hog cholera virus, gD protein of porcine pseudorabies virus, gB protein of porcine pseudorabies virus and cap protein of porcine circovirus type 2. 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 infection of swine erysipelas filamentous bacillus can be effectively prevented and protected.

The present disclosure provides a recombinant E2 protein of classical swine fever virus (CSFV) comprising an amino acid sequence as set forth in SEQ ID NO: 2, or an amino acid sequence having at least 85% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2. The recombinant E2 protein of the present disclosure can comprise a point mutation at one or more of amino acids at positions 48, 63, 77, 81, 86, 95, 103, and 129. The present disclosure also provides a nucleic acid molecule encoding the recombinant E2 protein, a host cell comprising the nucleic acid molecule, an immunogenic composition comprising the recombinant E2 protein, and uses thereof.

The application discloses a recombinant pseudorabies virus strain expressing classical swine fever virus (CSFV) recombinant E2 protein and application, and relates to the technical field of biology.The TK / gE double gene deletion PRV attenuated strain is used as a virus carrier, an exogenous CSFV E2 gene with a transmembrane region removed is inserted after a PRV gC signal peptide sequence, and a recombinant virus rPRV-delTK / gE-gC-E2 for expressing the gC N terminal and fusing with the PRV gC is constructed; Western blotting analysis and indirect immunofluorescence detection prove that the gC-E2 protein is successfully expressed.The recombinant virus immunized mice can produce specific antibodies against E2; after immunizing piglets, the piglets can produce an antibody level equivalent to that of a classical CSFV attenuated vaccine strain C strain.The rPRV-delTK / gE-gC-E2 obtained in the research is a promising bivalent live vaccine candidate strain.

The application provides a fusion protein and a double subunit vaccine simultaneously expressing BVDV E2 protein and BRSV F protein, and relates to the technical field of animal biological products.The fusion protein is subjected to enzymatic hydrolysis and in-vitro refolding to obtain a bovine viral diarrhea and bovine respiratory syncytial virus double subunit vaccine with complete structure and function, and the bovine viral diarrhea virus E2 protein is in the form of a dimer, and the bovine respiratory syncytial virus F protein is in the form of a trimer to form the vaccine.The application realizes the production of two kinds of virus subunit vaccines simultaneously in one expression process, the purification filler and the enzyme for enzymatic hydrolysis can be reused, the purification process is simple, the production cost is low, the antigen purity is high, no ADE effect is generated, and the immune side reaction is effectively reduced.

The present application discloses a chimeric classical swine fever virus, a classical swine fever marker vaccine, and a preparation method and application thereof. In the embodiment of the present application, the non-coding region and the main antigenic region of the E2 protein of the attenuated live classical swine fever vaccine strain C were replaced with the corresponding region of the BVDV Hubei strain by reverse genetic manipulation technology, and an infectious clone of the chimeric classical swine fever virus was constructed. The infectious clone was rescued by cell transfection and passage to obtain a chimeric classical swine fever virus with stable characteristics. The chimeric classical swine fever virus can stably replicate in pig-derived cells and has genetic stability and safety; it can induce the production of specific antibodies after immunization of the body, and can also provide the pig with complete protection against lethal infection by the virulent classical swine fever virus. In addition, the strain can immunize to produce BVDV E2-specific antibodies to distinguish between vaccinated pigs and wild-type classical swine fever virus-infected pigs, so that the strain has an application prospect as a classical swine fever marker vaccine.

The application discloses a recombinant pseudorabies virus strain expressing a full-length E2 protein of a mutant of a classical swine fever virus and application, and relates to the technical field of biology.The recombinant full-length E2 protein provided by the application has a hydrophobicity score lower than 3.25, and has a high protein expression level; meanwhile, the transmembrane region of the E2 protein has a hydrophobicity threshold value of 3.25-3.51.The finding has guiding significance for expression of the E2 protein or other transmembrane proteins in eukaryotic cells.The application constructs a recombinant PRV expressing a classical swine fever virus (CSFV) E2 protein, and high-level E2 antibodies and cellular immunity can be induced after immunization of mice; after immunization of rabbits, the rabbits can resist lethal attack of PRV ZJ2013 and fever reaction caused by the classical swine fever.The recombinant pseudorabies virus strain provided by the application can be used as a double vaccine for preventing CSFV and PRV infection.

The invention relates to the technical field of biological gene expression, in particular to a signal peptide for increasing protein secretion in a protein expression system and application of the signal peptide. The amino acid sequence of the signal peptide Hybrid for increasing the protein secretion in the expression system is as shown in SEQ ID NO. 4. The signal peptide Hybrid can remarkably improve the secretory expression quantity of hog cholera E2 protein or VP7 protein from porcine rotavirus, is excellent in secretory capacity, and can be used as signal peptide selection of secretory protein in different protein expression systems.

The invention discloses a recombinant pseudorabies virus strain for expressing hog cholera virus recombinant E2 protein and application, and relates to the technical field of biology. According to the research, a TK / gE double-gene deleted PRV attenuated strain is used as a virus vector, and after an exogenous CSFV E2 gene without a transmembrane region is inserted into a PRV gG signal peptide sequence, a recombinant virus rPRV-delTK / gE-gG-E2 which is positioned at a gG N end and is subjected to fusion expression with PRV gG is constructed; western blotting analysis and indirect immunofluorescence detection prove that the gG-E2 protein is successfully expressed. A recombinant virus immunized mouse can generate a specific antibody aiming at E2; after piglets are immunized, an antibody level equivalent to that of a classical CSFV attenuated vaccine strain C strain can be generated. The rPRV-delTK / gE-gG-E2 obtained by the research is a bigeminy live vaccine candidate strain with an extremely promising prospect.

According to the invention, swine antiviral proteins IFITM1, IFITM3 and Viperin are used as immunopotentiators and are applied to the swine fever virus E2 protein antiviral baculovirus inactivated vaccine. When swine antiviral proteins IFITM1, IFITM3 and Viperin serve as immunopotentiators and are used for immunization together with the swine fever virus E2 protein antiviral baculovirus inactivated vaccine, immunized mice can be promoted to generate higher-content swine fever virus serum IgG antibodies. At present, the inactivated vaccine of the antiviral protein immunopotentiator for the pigs is not put into use in the domestic market. However, the swine immunopotentiator adopted in the scheme is matched with the swine fever virus E2 protein antiviral baculovirus inactivated vaccine, so that the swine fever can be more effectively prevented and controlled, and the blank in the aspect of immunopotentiators in the current pig raising market is filled.

The invention relates to the technical field of antibodies, in particular to an anti-Getavirus monoclonal antibody 1D8, a nucleic acid molecule and application of the anti-Getavirus monoclonal antibody 1D8. The monoclonal antibody 1D8 secreted by the hybridoma cell strain 1D8 provided by the invention can be subjected to specific immunoreaction with GETV and GEV-E2 proteins, has good immunogenicity, good specificity and high sensitivity, can be widely applied to an immunodetection technology, can be used as an antibody for Western Blot and indirect immunofluorescence tests, and is used for identifying GETV and E2 proteins and measuring the content of GETV viruses; meanwhile, the antibody is obtained by using a conventional gene engineering or protein engineering method, so that the long-term cryopreservation antibody gene loss of hybridoma cells is avoided, the optimization of the antibody on gene and protein levels is also facilitated, and the specificity and affinity of the antibody are further improved.

This invention provides a mutant human papillomavirus (HPV) type 16 E2 protein and its application. The mutant E2 protein is obtained by the following three mutations based on the wild-type HPV type 16 E2 protein: (1) the serine at position 317 of the wild-type HPV type 16 E2 protein is mutated to proline; (2) the threonine at position 17 of the wild-type HPV type 16 E2 protein is mutated to arginine, and the asparagine at position 55 is mutated to arginine; (3) the histidine at position 218 of the wild-type HPV type 16 E2 protein is mutated to proline. This invention obtains the target protein through amino acid mutation and codon optimization, resulting in a more stable protein structure that is more conducive to expression, thereby significantly improving the yield and preparation efficiency of the HPV type 16 E2 protein. This antigen can be used for the detection of HPV type 16 E2 antibodies or to evaluate the humoral immunogenicity of therapeutic vaccines targeting HPV type 16 E2.

Provided are a fusion antigen of porcine Getah virus (GETV), a kit, a preparation method therefor and an application thereof. The fusion antigen of the GETV is primarily prepared by recombining a Gaussia luciferase (GLuc) gene with a codon- optimized GETV E2 antigen gene to construct an expression vector, and transfecting the expression vector containing GLuc-E2 into mammalian cell lines, resulting in the secretion of GLuc-E2 proteins into a cell supernatant for expression. Without the need for protein purification step, the cell supernatant may be directly collected for disease detection. The present disclosure demonstrates strong specificity and shows no cross- reactivity with African swine fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV), or Japanese encephalitis virus (JEV).

The invention is a composition for the treatment of HPV infections, which, with its components containing [3-caryophyllene, geranyl acetate, geraniol, and nerol, modulates the E7 and E2 protein pathways to provide a natural supportive therapeutic effect in managing HPV infections, exhibits antiviral, antibacterial, and immunomodulatory activity, significantly reduces viral replication, and suppresses protein expression, Contains Eugenia aromaticum (clove oil), which prevents viral replication by disrupting the structural integrity of cell membranes, exhibiting antimicrobial and antiviral activity with its high eugenol content, and Contains Cymbopogon martinii (palmarosa oil), which disrupts viral cell walls and inhibits the replication process, exhibiting antiviral and antibacterial properties with its high geraniol and citronellal content, and Contains Melaleuca viridiflora (niaouli oil), which strengthens cellular immune mechanisms to limit the intracellular spread of viruses and exhibits antiviral and immunomodulatory effects with its viridiflorol content.

The application discloses a CSFV-PPV 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 shown in SEQ ID NO:1 or an increased or reduced sequence thereof. The second gene has a sequence shown in SEQ ID NO:2 or an increased or reduced sequence thereof. The application takes the antigen E2 protein of CSFV (CSFV) and the VP2 protein of PPV (PPV) as double targets, expresses the recombinant SC-E2 protein with a SpyCatcher label and the recombinant ST-VP2 protein with a SpyTag label in insect cells through a recombinant baculovirus vector, realizes in-vitro covalent assembly of the double antigens, and constructs the bivalent subunit vaccine which can simultaneously prevent and control two diseases, and has the advantages of high safety, strong immunogenicity, high prevention and control efficiency and easiness in large-scale production.

The application provides a bivalent bovine viral diarrhea virus antigen protein composition, a subunit vaccine and a preparation method and application thereof, and belongs to the technical field of genetic engineering vaccines.The bivalent bovine viral diarrhea virus antigen protein composition is composed of a recombinant protein kozak-MLTsp-BVDV1-E2-His and a recombinant protein kozak-IgKsp-BVDV2-E2-His, the amino acid sequence of the recombinant protein kozak-MLTsp-BVDV1-E2-His is shown as SEQ ID NO.16, and the amino acid sequence of the recombinant protein kozak-IgKsp-BVDV2-E2-His is shown as SEQ ID NO.18.The application realizes efficient expression of BVDV-1 and BVDV-2 E2 proteins, obtains high-purity E2 proteins, ensures the safety and effectiveness of the vaccine, and improves the antibody level of the vaccine.

The application discloses a recombinant 2.1d subtype swine fever E2 protein, a subunit vaccine and application thereof, and belongs to the technical field of genetic engineering. By removing amino acids at positions 344-375 of the 2.1d subtype swine fever E2 protein and retaining amino acids at positions 1-343, the most important antigen epitope of the 2.1d subtype swine fever E2 protein having a protective effect can be retained, the spatial structure of the recombinant 2.1d subtype swine fever E2 protein is basically not affected, and the immunogenicity of the recombinant 2.1d subtype swine fever E2 protein can be retained to the maximum extent. In addition, the recombinant 2.1d subtype swine fever E2 protein can be efficiently expressed in a prokaryotic expression system. Further, by co-expressing a molecular chaperone, the soluble expression level of the recombinant 2.1d subtype swine fever E2 protein can be significantly improved. After the recombinant 2.1d subtype swine fever E2 protein is prepared into a subunit vaccine, the subunit vaccine has good protective efficacy on infection of a 2.1d subtype swine fever virus.