45 results about "HN Protein" patented technology

The invention relates to an epitope peptide H362 of an HN protein in PPRV, and determination, a preparation method and application thereof. The amino acid sequence of the epitope peptide is H362: <362>EANWVVPSTDVRDL<375>. The invention detects reactogenicity of a monoclonal antibody and PPRV and specificity of the monoclonal antibody; according to detection results, the monoclonal antibody has good reactogenicity to rPPRV-HN-F protein; immunoinformatic technology is cooperatively used for predicating the B cell epitope of the HN protein; an aminated ELISA plate is employed for detecting candidate epitopes and the monoclonal antibody 10E3, and the epitope peptide H362 corresponding to 10E3 is determined; and determination of the epitope peptide lays a theoretical foundation for preparation of epitope vaccine antigens and diagnostic reagent antigens for PPRV.

The invention relates to a preparation method and application of fusion protein (chIL-2-HN fusion protein) of recombining poultry interleukin-2 (IL-2) and newcastle disease virus hemagglutinin-neuraminidase (HN), belonging to the technical field of biological engineering. The fusion protein is fused by poultry IL-2 protein and newcastle disease virus HN protein by flexible Linker peptide; a DNA sequence for coding the fusion protein is inserted into an expression vector pPICZ alpha A; saccharomycete X-33 is electrically converted to obtain a genetically engineered microorganism for efficiently expressing recombination chIL-2-HN fusion protein which is prepared by liquid culture and purification; the recombination fusion protein can serve as the novel genetic engineering vaccine and can serve as a novel immunologic adjuvant for newcastle disease common vaccine. The chIL-2-HN fusion protein of the invention has favourable safety and no toxic or side effect, which is verified by experiments of animal immunoassay. In addition, the chIL-2-HN fusion protein can effectively strengthen the level of organism cellular immunity and humoral immunity and has wide application prospect.

The invention relates to the technical field of Newcastle disease vaccine in particular to Newcastle disease virus-like particles, a preparation method and a patent application of an application thereof. The virus-like particles are protein particles with a hollow structure formed by the self-assembly of Newcastle disease virus M protein, F protein, NP protein and HN protein. The base sequence of a M1 gene corresponding to the M protein is shown in SEQ ID NO.1, the base sequence of the F1 gene corresponding to the F protein is shown in SEQ ID NO.2, the base sequence of the NP1 gene corresponding to the NP protein is shown in SEQ ID NO.3, the base sequence of the HN1 gene corresponding to the HN protein is shown in SEQ ID NO. 4. The Newcastle disease virus-like particles, preparation method and application have the technical advantages of high expression level of structural protein, self-assembly of the virus-like particles, immunogenicity closer to real virus particles, high yield and purity of the virus-like particles, and good immunization effect.

The invention relates to establishment of a hybridoma cell strain for secreting duck NDV (newcastle disease virus)-resisting isolate monoclonal antibody, which belongs to the technical field of molecular immunology and virology. The purified HN protein expressed by duck NDV isolate SDO3 pronucleus is used as immunogen, one hybridoma cell strain capable of secreting duck NDV-resisting isolate monoclonal antibody is researched, and the preservation number is CCTCC C2013173. The NDV specificity monoclonal antibody secreted by the hybridoma cell strain cannot be specially combined with the NDV strain but can be specifically combined with NDV clinical wild strain, so that the NDV specificity monoclonal antibody can be used as an identification reagent to be used for identifying the NDV vaccine virus and clinical wild strain, and the specificity is strong; the sensitivity is 1: 212 HAU; the hybridoma cell strain has the advantage of high sensitivity.

The invention provides a human parainfluenza virus quantum dot immunochromatography typing detection card, a preparation method and applications. The detection card comprises a base plate, a sample pad, a water absorption pad, a combination pad and a detection layer. The combination pad is coated with a mixture of rabit-anti I-type, II-type and III-type human parainfluenza virus HN protein polyclonal antibodies labeled with quantum dots respectively. The detection layer is composed of a solid phase nitrocellulose membrane with three detection lines and a quality control line. The three detection lines are coated with rabit-anti I-type, II-type and III-type human parainfluenza virus polyclonal antibodies respectively. The quality control line is coated with anti-rabit IgG. The detection layer is pasted on the base plate. The combination pad and the water absorption pad are arranged above two ends of the detection layer respectively, overlap with part of the detection layer and are pasted with the detection layer and the base plate respectively. The sample pad is arranged on the combination pad, overlaps with part of the combination pad and is pasted with the combination pad and the base plate. The provided detection card has advantages of simple operation, rapid detection, quantification, high sensitivity and the like.

The invention discloses an aptamer realizing specific binding with the newcastle disease virus as well as a screening method and applications of the aptamer. The aptamer is specific aptamer B53 obtained through screening by adopting the SELEX technology. Specifically, the random single stranded DNA library and primers are constructed and synthesized in vitro, the newcastle disease HN protein and the newcastle disease virus GM strain realizing prokaryotic expression are adopted as target molecules, after screening, PCR amplification, positive screening and inverse screening, the screened single stranded DNA library realizing specific binding with the newcastle disease virus GM strain is subjected to clone sequencing, and thus the specific aptamer B53 is obtained. The aptamer B53 has the capacity of inhibiting virus replication, virus blood clotting activity and plaque formation, can specifically recognize the newcastle disease virus HN protein and the newcastle disease virus GM strain, has no reaction to avian influenza virus, infectious bursal disease virus and SPF chick embryo allantoic fluid, and can be used for detecting the newcastle disease virus.

The present invention relates to an epitope of HN protein in Newcastle disease virus which can be recognized by an avian immune system and an antibody against the epitope, a method for detecting a Newcastle disease virus by using the antibody, and an antigenic variant of Newcastle disease virus carrying changes in the epitope. The epitope of HN protein and the antigenic variant of Newcastle disease virus can be used for developing efficient vaccines, and further, in diagnosing the Newcastle disease virus rapidly and exactly.

The invention relates to peste des petits ruminant virus HN protein epitope peptide. The amino acid sequences of the epitope peptide are as follows: H123: <123>KFLNPDREYDFRDLR<137>, or/and H185: <185>GTGCLGRTVTRA<196>, or/and H487: <487>IRGPRGRCH<495>, or/and H569: <569>ECFPWYHKVWCYHDCLI<585>. B cell epitopes of a target protein are predicted by virtue of multiple immunoinformatic software, the different predicted epitopes are respectively artificially synthesized, the reactogenicity of the predicted epitopes is verified by virtue of an indirect ELISA method, an aminated ELISA Plate is coated with different polypeptides, and the reactogenicity with an antibody of HN protein is detected, and therefore, the B cell epitopes of the PPRV HN protein are authenticated.

The invention belongs to the technical field of animal vaccine preparation, and particularly relates to a diad vaccine regarding HN protein and chicken alpha interferon as effective components, and a preparation method thereof. The diad vaccine is prepared by transfection of BHK-21 cells and fully expressing obtained target protein by using HN genes and chicken alpha interferon genes through the utilization of a genetic engineering technology, and mixing the target protein with an adjuvant. When the diad vaccine prepared through the utilization of the expression proteins of the HN genes and the chicken alpha interferon genes is injected in a chicken body, the HN proteins and the alpha interferon proteins play respective biological roles in the chicken body, so that the effects of positively preventing and treating other virus infections are achieved while the virus infection of the newcastle disease is prevented. The vaccine is totally safe and reliable, is capable of inducing an organism to generate specific antibodies continuously for a long time and improving gallinaceous immunity, and well overcomes the disadvantage that the traditional vaccine only can prevent one disease.

The invention relates to a preparation method and application of a recombinant chicken interleukin-18 (IL-18) and Newcastle disease virus hemagglutinin-neuraminidase (HN) fusion protein (chIL-18-HN fusion protein). The fusion protein is formed by fusing chicken IL-18 (chiIL-18) protein and Newcastle disease virus HN protein through a flexible Linker peptide; a DNA (Deoxyribonucleic Acid) sequence for coding the fusion protein is inserted into an expression vector pPICZalphaA and yeast X-33 is electrically converted to obtain genetic engineering bacteria for efficiently expressing the recombinant chIL-18-HN fusion protein; the recombinant chIL-18-HN fusion protein is prepared through liquid culture and purification; and the recombinant fusion protein can be used as a Newcastle disease novel genetic engineering vaccine and can also be used as a novel immunologic adjuvant for the Newcastle disease conventional vaccine. Animal immunologic tests prove that the chIL-18-HN fusion protein provided by the invention has the advantages of high safety, no toxic or side effect, capability of effectively enhancing cellular immunity and humoral immunity level of organisms and broad application prospect.

The invention provides a Newcastle disease virus gene type VI vaccine strain which is constructed by carrying out attenuated mutation on an F gene of a pigeon-derived gene type VI Newcastle disease virus, replacing the F gene of a Newcastle disease virus strain with the preservation number of CCTCC NO: V201968 and then carrying out genetic rescue. The amino acids at the 340 site, the 342 site, the 347 site and the 353 site of the HN protein of the Newcastle disease virus strain with the preservation number of CCTCC NO: V201968 are mutated into histidine, asparagine, lysine and arginine respectively. The invention also provides an application of the Newcastle disease virus gene VI type attenuated strain NDV-VIb in preparation of vaccines. The skeleton virus of the constructed gene VI type Newcastle disease virus attenuated strain comes from the modified gene VII type Newcastle disease virus attenuated strain, is safe and reliable, and reduces the risk of virulence reversion; the constructed gene VI type recombinant Newcastle disease virus attenuated strain is mainly used for preventing and controlling the Newcastle disease virus of pigeons, and the F gene is derived from host pigeons and has better immunogenicity than the gene VI type Newcastle disease virus strain derived from chicken.

The invention provides a vaccine composition. The vaccine composition comprises a dog parainfluenza virus antigen with the immunity effective dose, a dog bordetella bronchiseptica antigen with the immunity effective dose, and an adjuvant, wherein the dog parainfluenza virus antigen is selected from an inactivated dog parainfluenza virus antigen or at least one of F protein, HN protein, M protein, NP protein, P protein and L protein, and the bordetella bronchiseptica antigen is selected from inactivated dog bordetella bronchiseptica antigen or p68 protein. Compared with the vaccines in the prior art, the vaccine composition provided by the invention has the better immune effect, and has no side effects, and thus the risk that a veterinarian worker or a contactor is infected is reduced. The invention further relates to a kit. The kit contains the inactivated dog parainfluenza virus antigen or the subunit antigen, the attenuated live dog bordetella bronchiseptica antigen or the attenuated live dog parainfluenza virus antigen, and the inactivated dog bordetella bronchiseptica antigen or p68 protein.

The invention relates to a human parainfluenza virus type 3 (HPIV-3) wild strain and an application thereof. The preservation number of the wild strain is CCTCC NO:V201911, the wild strain has HN protein and F protein of an HPIV-3 virus, and the wild strain can be applied to an HPIV-3 virus infected mouse model and an HPIV-3 neutralizing antibody detection experiment, and can be applied to preparation of vaccines for preventing parainfluenza type 3. The human parainfluenza virus type 3 wild strain disclosed by the invention is a cloned purified high-titer HPIV3LZ1728C19 virus strain free fromspecific exogenous factor plaque formation, and a human parainfluenza virus type 3 HN protein subunit vaccine prepared from the virus can effectively prevent parainfluenza virus type 3 infection.

The application provides an immune composition and a genetically engineered subunit protein vaccine. The vaccine includes newcastle disease virus HN protein encoded by the nucleic acid molecule shownin SEQIDNO:1 or nucleic acid molecule with the nucleic acid sequence has more than 95% similarity with that of the nucleic acid molecule shown in SEQIDNO:1, and newcastle disease virus F protein encoded by the nucleic acid molecule shown in SEQIDNO:3 or nucleic acid molecule with the nucleic acid sequence has more than 95% similarity with that of the nucleic acid molecule shown in SEQIDNO:3. The vaccine adopts CHO cell eukaryotic expression, protein glycosylation is thorough, the antigen protein immunogenicity is good, the expression quantity is very high and up to (2-3) g/L, recombinant cellscan be subjected to large-scale suspension culture, the complexity of vaccine preparation is reduced greatly and the production cost is reduced.

The invention discloses a mumps virus HN antigen capable of detecting a mumps-resisting virus antibody, especially a mumps-resisting virus neutralized antibody and further discloses application of the antigen in preparation of detection agent for detecting the mumps-resisting virus neutralized antibody and a corresponding detection kit.

The invention discloses polypeptide and ELISA detection reagent kit for detecting a Newcastle disease virus antibody. The reagent kit is used for coating an elisa plate through screened Newcastle disease HN protein polypeptide. Through detection of combining capacity of a serum antibody and polypeptide in samples, and through assistance with the antibody of goat anti-chicken Ig gamma labelled with horseradish peroxidase, the level of Newcastle disease virus antibody is detected. The reagent kit is used for detecting avian influenza virus H5, H7, H9 subtype antibodies, chicken infectious Fabricius' bursal disease virus antibodies, and chicken infectious bronchitis virus antibodies to be negative. The reagent kit can detect that the hemagglutination inhibition (HI) titer is greater than or equal to 4log<2> serum, and can meet clinical serum sensitivity detection requirements. When the reagent kit and a hemagglutination inhibition method are both used for detecting clinical serum, the coincidence rate is as high as 98.7%. The reagent kit is simple to operate, free from special material requirements, good in result judgment objectivity, free from influence by subjective judgment by operators, and suitable for being used as a tool for detecting and monitoring the level of the antibodies after immunization with a Newcastle disease vaccine.

The invention discloses a recombinant baculovirus for expressing heat-resistant HN protein of the Newcastle disease virus as well as a preparation method and application of the recombinant baculovirus. The virus strain is preserved in China Center for Type Culture Collection, the preservation address is Wuhan University, Wuhan, China, and the preservation number is CCTCC NO: V202044. The preparation method of the ecombinant baculovirus comprises the following steps of carrying out point mutation on an HN gene of a Newcastle disease virus non-heat-resistant LaSota strain, including G49E, R269S,G293E, R353Q and G494D, inserting the mutated HN gene into a baculovirus genome, and carrying out virus rescue to obtain the recombinant baculovirus rAC-HN-N5 strain. The virus strain can efficientlyexpress mutant HN protein. Compared with wild-type HN protein of the LaSota strain, the mutant HN protein has the advantage that the thermal stability is obviously enhanced. The mutant HN protein expressed and purified by utilizing the recombinant baculovirus rAC-HN-N5 strain can be used for preparing a heat-resistant subunit vaccine for the Newcastle disease, so that the dependence of the vaccine on a cold chain is reduced, and a new thought is provided for the development of the subunit vaccine with high heat stability.

The invention discloses a heat-resistant Newcastle disease virus mutant strain and a preparation method and application thereof. The mutant strain is deposited in the China Center for Type Culture Collection, and the deposit address is Wuhan University, Wuhan, China, and the deposit number is CCTCC NO: V202041. The preparation method uses the Newcastle disease virus TS09-C strain as the parent strain, and introduces 4 amino acid mutations into its HN protein: the 3rd arginine is mutated to serine, and the 197th arginine is mutated to isoleucine. acid, histidine at position 203 was mutated to asparagine and lysine at position 495 was mutated to valine. The heat resistance test results show that the heat resistance of the virus mutant strain is significantly higher than that of the parent strain, and is much higher than that of the commonly used vaccine strains, which proves that the present invention further improves the thermal stability of the existing heat-resistant strains.

The invention relates to the field of biotechnology, in particular, it provides a Newcastle disease virus vaccine. Prepare the protein of Newcastle disease virus vaccine, including the F protein of Newcastle disease virus and the HN protein of Newcastle disease virus, wherein, the nucleotide sequence of encoding F protein is as SEQ ID NO.1 (gene type VII) or SEQ ID NO.3 ( Gene Type II), the nucleotide sequence encoding HN protein is shown in SEQ ID NO.2 (Gene Type II) or SEQ ID NO.4 (Gene Type VII). The protein is suitable for the eukaryotic cell expression system, simplifies the process for preparing the active ingredient of the Newcastle disease virus vaccine, and reduces the cost at the same time. Moreover, the protein has broad-spectrum antigenicity and can achieve a good cross-protection effect.

The invention provides a gene VII type Newcastle disease virus vaccine strain, which is a gene VII type Newcastle disease virus attenuated strain NDV-VIIb strain, and is prepared by respectively mutating 340-site amino acid, 342-site amino acid, 347-site amino acid and 353-site amino acid of HN protein of a Newcastle disease virus strain with the preservation number of CCTCC NO: V201968 into histidine, asparagine, lysine and arginine; and then constructing through genetic rescue. The gene VII type Newcastle disease NDV-VIIb strain provided by the invention is used for preparing vaccines. The immune effect of the Newcastle disease virus gene VII type attenuated strain provided by the invention is obviously superior to that of a parent strain, and a good protection effect can be provided for the current popular gene VII type Newcastle disease virus.

The invention provides an HN protein mutant gene VII type Newcastle disease virus recombinant vaccine strain. A gene segment used in the process of constructing the gene VII type Newcastle disease virus recombinant vaccine strain contains a envelope glycoprotein F protein gene and an HN protein gene of a gene VII.2 subtype Newcastle disease virus with weakened toxicity. The gene VII type Newcastle disease virus recombinant vaccine strain provided by the invention has the biological characteristics of high growth titer and low pathogenicity in a chicken embryo, is stable in heredity, has a good immune protection effect on the Newcastle disease virus, can effectively inhibit toxin expelling, can be used for preventing and controlling the currently popular gene VII type Newcastle disease virus, and has a wide application prospect.