118 results about "Vaccine research" patented technology

Monoclonal antibody reagents that recognize the SARS-coronavirus (SARS-HCoV) are needed urgently. In this report we describe the development and immunochemical characterisation of mAbs against the SARS-HCoV based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of seventeen mAbs. Five mAbs exhibited Western immunoblot reactivity with the denatured spike protein, of which two demonstrated the ability to neutralize SARS-HCoV in vitro. Another four Western immunoblot-negative mAbs also neutralize the virus. These antibodies will be useful for the development of diagnostic tests, pathogenicity and vaccine studies.

The invention discloses human adenovirus antigen epitope chimeric protein as well as preparation and application thereof, and relates to fields of gene engineering techniques, vaccines and diagnostic reagents. The amino acid sequences of hexon protein of type 3, type 7, type 11, type 14 and type 55 of adenovirus are analyzed through computer analysis, protein fragments with good antigenicity can be screened respectively, the fragments are connected by using two glycine and one serine, then chimeric protein with multiple antigen fragments in serial connection can be formed, pronucleus preferred codons are selected and interpreted into corresponding nucleotide sequences, and full-length genes can be synthesized in a chemical manner. The chimeric protein can be obtained through expression purification by using a gene engineering technique, and the chimeric protein comprises 363 amino acids in whole length. The expressed chimeric protein can be applied to vaccine research, HAdV antibody or antigen detection and the like, and is related to fields such as gene engineering techniques, vaccines and diagnostic reagents.

The invention relates to the technical field of swine pseudorabies viruses, and relates to a swine pseudorabies virus variant XF-1 strain, a preparing method thereof and applications of the strain. The accession number of the variant is CCTCC NO V201654. The swine pseudorabies virus variant (the XF-1 strain) is adopted to prepare an inactivated vaccine, and safety and immunoprotective experiments are performed. Results prove that the swine pseudorabies virus variant (the XF-1 strain) inactivated vaccine has good safety, and that the immunoprotection efficiency of the inactivated vaccine is obviously higher than that of other immune groups, and therefore the virus strain is proved to have good immunogenicity, can be used for vaccine research and development for the swine pseudorabies, and has a food vaccine development prospect.

The invention provides a cell culture method for duck flavivirus. The method comprises the following steps: inoculating to attach an anchorage-dependent kidney cell BHK21 of a baby hamster to the surface of a cell culture container to form a monolayer; inoculating the monolayer with virus; culturing the monolayer inoculated with the virus in an incubator with 5% CO2 at 37 DEG C; and continuously passing by taking a cell culture product as an inoculum for the next passage. According to the cell culture method provided by the invention, after the virus is continuously passed for 5-7 generations on the monolayer, stable multiplication can be obtained; and after the cell is inoculated for 3-4 days, an obvious cytopathic effect can be formed. The method provides a convenient and visual virus operating platform for studying the biological characteristics of the virus and the molecular basis of virus pathopoiesia and for the vaccine research developed for effectively preventing and controlling the disease.

The invention discloses a respiratory syncytial virosome vaccine, which mainly comprises a respiratory syncytial virosome, wherein the virosome comprises a complete respiratory syncytial virus (RSV) surface, and can stimulate good dual cell and body fluid immune response. The virosome protein antigens are prepared into formulations such as injection, nasal spray, sublingual tablet and transdermal agent by adding or not adding an adjuvant, can safely and effectively prevent RSV infection after being immunologically inoculated to different animals or people, and provide the ideal vaccine for the safe and effective immune prevention and control of the RSV infection of different age groups. Researches show that the respiratory syncytial virosome vaccine has a good immunological protection effect on the different age groups.

The invention discloses a cattle food-and-mouth disease virus A type synthetic peptide. The cattle food-and-mouth disease virus A type synthetic peptide has the following amino acid sequence: acetyl-YDLDF EALKP HFKSL GQTIT PADKS PPS VYNGT CKYSA PATRR GDLGS LAARL AACLP ASFNY GAIRA T-amide. The cattle food-and-mouth disease virus A type synthetic peptide can be used as an effective novel vaccine for the A type food-and-mouth disease in production practice; simultaneously, the realization of the test provides a certain foundation for further perfecting the construction of a food-and-mouth disease virus A type synthetic peptide vaccine and the construction of other food-and-mouth disease subtype synthetic peptide vaccines; and the new ideal proposed in the research of the synthetic peptide vaccine and the construction of a new method of the synthetic peptide vaccine provide theoretical foundation and technical support for further perfecting the development and research of the multiple antigenic peptide, multivalent peptides and joint peptide vaccines in future.

The invention discloses a pig epidemic diarrhea attenuated strain as well as a culture method and application thereof. The preservation number of the pig epidemic diarrhea attenuated strain JS / PEDV / 2 / 2014G100 is CCTCC NO.V201728; the whole length of the strain is 27939bp; 49 nucleotides are deleted from the ORF3 gene of the strain when being compared with a PEDV virulent strain; phylogenetic analysis shows that the strain belongs to a G1 subgroup, can be proliferated without pancreatin on Vero cells, belongs to cell adaptation strains and is relatively good in cross protection, antigen property and immunogenicity; pigs infected by the strain have normal clinical symptoms, the weights of the pigs can be increased normally, no toxin removal phenomenon is caused, high-level PEDV antibodies can be generated in pig bodies, the OD450 value of a serum IgG antibody can be up to 1.2, the OD450 level of an immunized 60d antibody can be still maintained at 1.0, and scientific research materials are provided for later vaccine research and development.

The invention discloses a bovine A-type foot-and-mouth disease multi-epitope recombinant vaccine, and a preparation method and an application thereof, and belongs to the field of veterinary vaccine research. The preparation method comprises the following steps: carrying out reasonable serial connection on the dominant antigen epitopes of bovine A-type foot-and-mouth disease virus representative strains once pandemic in China by adopting an antigenized antibody strategy and a brand new reverse vaccinology technology and strategy, coupling with a bovine IgG immunostimulatory fragment (IgG heavy chain constant region), cloning into a prokaryotic expression vector to construct a recombinant expression vector, transforming Escherichia coli cells to express a recombinant antigen, purifying by adopting Ni-NAT column chromatography, quantifying by a Bio-Rad protein quantification kit, and preparing the vaccine individually or combining with a recombinant foot-and-mouth disease virus 3D protein. A result of animal immune experiments shows that the recombinant protein or combined vaccine can stimulate the body to produce a protective antibody with high titer and also can protect immune animals against a virus attack, so the bovine A-type foot-and-mouth disease multi-epitope recombinant vaccine has a good application prospect.

The invention relates to a 1 A1S-1969 recombinant protein and a preparation method and application thereof. The recombinant protein comprises 46th to 414th site amino acid sequence of 1 A1S-1969, and the amino acid sequence of the recombinant protein is shown as SEQ ID NO. 5. The recombinant protein is high in expression amount, easy to separate and purify, efficient and safe, can be directly used with an adjuvant, and is used for the preparation of an acinetobacter baumannii infection resistant subunit vaccine and a related detection kit. Confirmed by animal experiments, the genetic engineering recombinant subunit vaccine has good acinetobacter baumannii infection resistant immune protection effect, lays a foundation for the further study on combined vaccines and multicomponent fusion vaccines, and plays an important role for development and application of prevention and control vaccines and diagnostic kits.

The invention discloses a bovine A-type foot-and-mouth disease broad-spectrum multi-epitope recombinant vaccine, and a preparation method and an application thereof, and belongs to the field of veterinary vaccine research. The preparation method comprises the following steps: carrying out reasonable serial connection on the major antigen epitopes of pandemic in China and bovine A-type foot-and-mouth disease virus representative strains recently pandemic in countries bordering China by adopting a brand new reverse vaccinology idea and strategy, coupling with a bovine IgG heavy chain constant region, cloning into a prokaryotic expression vector to construct a recombinant expression vector, transforming Escherichia coli cells to express a recombinant antigen, purifying by adopting Ni-NAT column chromatography, quantifying by a Bio-Rad protein quantification kit, and preparing the vaccine individually or combining with a recombinant foot-and-mouth disease virus 3D protein. A result of animal immune experiments shows that the multi-epitope antigen or 3D protein combined vaccine can stimulate the body to produce a protective antibody with high titer and also can protect immune animals against a virus attack, so the bovine A-type foot-and-mouth disease broad-spectrum multi-epitope recombinant vaccine has a good application prospect.

The present invention belongs to the field of virus gene engineering, and relates to virus-like particle containing RNA virus nucleic acid and its preparation process and application. The virus-like particle is RNase resistant, and contains coat protein and inside recombinant gene sequence including partial MS2 bactereophage sequence, RNA virus nucleic acid sequence and partial RNA virus antigen. The virus-like particle of the present invention may be used as stable and reliable RNA quality controlling and standard article without biological infectious danger. The present invention provides also RT-PCR reagent kit amplification segment suitable for detecting virus RNA in several methods. The virus-like particle containing partial RNA virus antigen may be used as the mold for research of the interaction between virus and host cell, as antisense RNA tool for directional transportation of small molecule medicine, and in vaccine research.

The invention discloses a method for extracting and purifying eimeria tenella merozoite. The method comprises the steps of: releasing the merozoite and purifying the merozoite, wherein the merozoite release comprises mechanical grinding release and enzymic digestion release, 0.0625% pancreatin and 0.175% sodium taurodeoxylate are adopted as digestive juice in the enzymic digestion release process; digestion is moderate, which is beneficial to the activation of the merozoite, can eliminate erythrocytes to obtain pure merozoite. The method is convenient and simple to operate, is low in cost, and is ideal in separation and purification effects; the separated merozoites are large in quantity, are pure with few impurities and keep vigorous energy; and the method is suitable for research on drug susceptibility detection, vaccine research and other biological researches.

The invention provides a replicon vector taking Japanese encephalitis virus genome as a framework, as well as a cell line for packaging the same and a packaging system. The JEV replicon vector has the capability of efficiently expressing foreign protein. Mice are immunized with the replicon vector, and the titer of an anti-JEV antibody reaches 1:1280 after three immunizations so as to protect 75 percent of suckling mice from virus attack. The cell line provided can produce 1.6*105 U / ml of pseudovirus particles after packaging JEV replicon, and the titer of the anti-JEV antibody reaches 1:2560 after two immunizations so as to protect 73 percent of suckling mice from virus attack. The invention establishes a technical platform for a JEV replicon vector system for the first time, and explores the feasibility of researching replicon vaccines and pseudovirus vaccines through the JEV replicon vector system, thereby laying a solid foundation for developing and researching a plurality of novel vaccines used to prevent and treat tumors and viral diseases in future.

The invention discloses a pig acute communicable disease virus infectious DNA carrier preparation method and the application, this carrier includes T7 to start sub- and pig acute communicable disease virus standard full strength Shimen gene group's DNA, starts sub- and in downriver DNA in T7, in the flaw NS2 gene 1,260 alkali base to, retains the 5' end 3752nd alkali base to to the PshAI position spot 3881st alkali base to the between 130bp NS2 gene fragment, other DNA sequence and a Shimen infectiousness clone DNA to be same. The invention method is simple, the ease of operation, may construct the double price or the multi- prices genetic engineering vaccine, suits the pig acute communicable disease virus the distinction diagnosis and in the vaccine research application.

The invention provides a new coronavirus SARS-CoV-2 broad-spectrum polypeptide antigen, a specific neutralizing antibody thereof and application, and belongs to the technical field of virus immunodetection. The new coronavirus SARS-CoV-2 broad-spectrum polypeptide antigen has an amino acid sequence shown as SEQ ID NO: 1, and can be specifically combined with a new coronavirus antibody through reaction with SARS-CoV-2 human positive serum. Based on the polypeptide sequence, triple SARS-CoV-2 broad-spectrum polypeptide tandem fusion protein is prepared by utilizing PCR, prokaryotic expression and protein purification technologies, a trimer mode of SARS-CoV-2S protein in a natural state is simulated, the fusion protein is used as an antigen to immunize mice, the SARS-CoV-2 specific neutralizing antibody can be generated, and the neutralizing antibody has good application prospects in SARS-CoV-2 anti-infection treatment, vaccine research and development and detection kit development.

The invention discloses a grass carp swim bladder epithelioid cells line and an application. According to the grass carp swim bladder epithelioid cells line with a preservation number of CCTCC No.C201443, Grass Carp Reovirus GCRV propagation amount in the cell line is obviously higher than that of the cell lines such as CIK, CO, PSF, FHM and EPC used for separating GCRV, cell pathology time is fast, and the cell line can better used in the researches of separation and breeding of Grass Carp Reovirus and grass carp hemorrhagic disease vaccine.

The invention relates to the technical field of biological medicines, and particularly discloses a method and a kit for detecting a new coronavirus IgG / IgM total antibody. According to the detection method, the IgG / IgM total antibody is detected by using a double-antigen sandwich technology. The detection method specifically comprises the following steps of coating a recombinant new coronavirus RBD protein on a solid-phase carrier, adding a to-be-detected sample into the solid-phase carrier coated with the recombinant new coronavirus RBD protein, incubating and washing, adding biotin-labeled new coronavirus RBD protein into the washed solid-phase carrier, incubating, washing, developing and combining with a standard curve to obtain the antibody content. The technical principle of double antigen sandwich is utilized to detect that the serum stock solution of a healthy person has no false positive, so that the healthy person and a patient can be well distinguished; the method can promote large-scale early screening emergency serological detection of the new coronavirus and related vaccine research and vaccine effect evaluation work.

The invention discloses a CA-193 virus strain (Coxackievirus A16) and application thereof to preparation of an inactivated vaccine. For the application, MRC-5 cells are used for proliferating the CA-193 virus strain; virus proliferation liquid is subjected to centrifugation, concentration and column chromatography filtration and purification after inactivation by formaldehyde or beta-propiolactone; and a CA16 inactivated vaccine is obtained. The invention provides the vaccine virus strain capable of being used for preparing a human CA16 inactivated vaccine; the subtype of the vaccine virus strain belongs to the main epidemiological subtype B1 in Chinese Mainland; the vaccine virus strain has good growth characteristics on the MCR-5 cells; and the virus titer (7.5 to 8.251gTCID50 / ml) with high stability can be obtained. A preparation method of the vaccine is complete; the impurity content is low; the immunogenicity and the immunizing protection performance are good; meanwhile, cell culture substrates are MRC-5 cells; the safety is high; and the international and CFDA specifications and requirements on the vaccine research and development can be well met.

The invention discloses a broad-spectrum multi-epitope recombinant vaccine for bovine foot-and-mouth disease virus type A strain epidemic abroad and a preparation method and application thereof, and belongs to the field of veterinary vaccine research. Adopting the ''antigenized antibody'' strategy and a novel reverse vaccine operation technique and strategy, the method is as below: conducting reasonable combination of the dominant antigen epitopes of representative strains of bovine foot-and-mouth disease virus type A recently epidemic in countries bordering with China; conducting coupling of the epitopes with bovine IgG heavy chain constant region; cloning into a prokaryotic expression vector to construct a recombinant expression vector; transforming into E.coli competent cell to express the recombinant antigen, and purifying by using an Ni-NAT chromatographic column; quantifying by a Bio-Rad protein quantification kit; preparing the vaccine by using the antigen individually or combining the antigen with a recombinant foot-and-mouth disease virus 3D protein. Animal immune experiment results show that the recombinant protein or vaccine after compatibility can stimulate the body to produce protective antibody with high potency and at the same time protect the immune animals against homologous virus attacks; therefore, the invention has good application prospect.

The invention discloses antigen epitope peptide of novel coronavirus T cells and application of the antigen epitope peptide. The amino acid sequence of the antigen epitope peptide is shown as any one of SEQ ID NO: 1-13. A pMHC compound monopolymer is prepared from the pMHC compound, a pMHC compound polymer is further prepared, and the pMHC compound monopolymer can be used for detection of antigen-specific T cells in peripheral blood of a novel coronavirus infected convalescent patient and used for in-vitro T cell activation experiments. The novel coronavirus T cell antigen epitope peptide can be applied to immunodetection and vaccine research and development related to novel coronavirus, and is worthy of deep research and vigorous popularization.

The invention discloses a porcine deltacoronavirus strain, which is assigned with the accession number of CCTCC NO:V201558. According to the obtained porcine deltacoronavirus strain, an important foundation is to be laid for the etiology investigation, epidemiological investigation, diagnosis prevention and control and vaccine research of the virus.

The invention discloses double RT-PCR (reverse transcription-polymerase chain reaction) primer, kit and method for amplifying North America and European porcine blue ear disease viruses; the method has the advantages of high specificity, high sensitivity, low time and labor consumption and the like, and is of important guidance significance to molecular epidemiology, genetic evolution, vaccine research and development and the like for studying porcine blue ear disease. In addition, denaturing and annealing temperature in the method only takes 20 s, amplification can be performed as well just with one PCR apparatus, and the method may also act as a method for identifying and detecting North America and European porcine blue ear disease viruses and helps quickly classify porcine blue ear viruses.

The invention provides a chimeric type porcine circovirus live vaccine C1-233 strain and a constructing method thereof, and belongs to the technical field of vaccine study production. The constructing method comprises the steps of screening of PCV1 and PCV2 strains, construction of a recombined porcine circovirus 1 type live vector vaccine strain which expresses a porcine circovirus 2b type ORF2 gene, recombination of viral nucleic acid, and composition characteristics and biological characteristics of an amino acid sequence. Finally a recombined porcine circovirus 1 type live vector vaccine strain (C1-233 strain) which can be used for industrialized production and expressing the porcine circovirus 2b type ORF2 gene is obtained.

The invention relates to an A1S-1462 recombinant protein and a preparation method and application thereof. The recombinant protein comprises A1S-1462 mature peptide, and the amino acid sequence of the recombinant protein is shown as SEQ ID NO. 3. The recombinant protein is high in expression amount, easy to separate and purify, efficient and safe, can be directly used with an adjuvant, and is used for the preparation of an acinetobacter baumannii infection resistant subunit vaccine and a related detection kit. Confirmed by animal experiments, the genetic engineering recombinant subunit vaccine has good acinetobacter baumannii infection resistant immune protection effect, lays a foundation for the further study on combined vaccines and multicomponent fusion vaccines, and plays an important role for development and application of prevention and control vaccines and diagnostic kits.

A hepatitis C virus (HCV) genome is highly heterogeneous, and can be divided into eight genotypes and multiple gene subtypes according to the difference of genomic bases. HCV from patients cannot be cultured directly in cells in vitro. Infectious clones of HCV for constructing epidemic strains by a reverse genetic method is an important technical operation platform for promoting HCV basic researchand drug and vaccine research and development. The gene type 6 HCV is mainly prevalent in China and Southeast Asia. At present, no full-length infectious clones of the gene type 6 Chinese strain exist. The main content comprises establishment of in-vitro cell infectious clones CH6acc of the HCV gene type 6a in China and demonstration of application of the CH6acc in drug research. A CH6acc cell culture model can be used in HCV basic research, and drug and vaccine research and development.

The invention provides a method for culturing myxobolus in vitro and belongs to the technical field of parasite in-vitro culture. Crucian muscle homogenate and crucian gill homogenate are adopted as culture carriers for myxobolus in-vitro culture. The method includes the following steps of firstly, preparation of antibiotics; secondly, preparation of a supernatant culture solution of the crucian gill tissue homogenate; thirdly, in-vitro culture of myxobolus; fourthly, counting and growth curve drawing. The culture conditions of myxobolus are optimized, myxobolus can propagate in a proper environment outside a host, a large number of base materials are provided for later pharmacology or vaccine research, and further research on diseases or myxosporean caused by myxobolus is promoted.

The invention provides a porcine circovirus type 3 strain PCV3-LY and application thereof. The virus is the porcine circovirus type 3 rescued by infectious cloning technology at home and abroad for the first time. After the virus is inoculated to monolayer porcine kidney cells PK15 for adherent culture, D-glucosamine treatment is carried out, then culture is continued, a cell culture is harvestedand used as an inoculum of a next round of passage, and finally a cell culture-adapted strain is obtained by successive passages. During the adaptation process, significant cytopathic changes can be observed after 15 passages. Indirect immunofluorescence confirms that the cell is able to support efficient replication of porcine circovirus type 3. The titer of the virus is lgTCID[50]=10<6.53> / mL. An animal regression test finds that two pigs are killed after five 28-day-old pigs are challenged with the virus, and the virus is more pathogenic to piglets. The virus is the porcine circovirus type3 rescued by infectious cloning technology at home and abroad for the first time, and provides direct support for further research on pathogenesis of PCV3 and development of vaccine research for effectively preventing and controlling the disease.

The invention discloses an Eimeria tenella rhoptry protein 41 as well as a preparation method and application thereof. The protein is a new-found protein capable of highly expressing in a sporozoite stage of Eimeria tenella and is a protein relevant with invasion of sporozoites in host cells, an amino acid sequence of the protein includes an amino acid sequence represented by SEQ ID NO.1. An EtROP41 gene is linked with an escherichia coli expression vector to form a recombinant expression vector, and a recombinant EtROP41 protein is expressed in escherichia coli, is purified and is mixed withan adjuvant so as to form a chicken Eimeria tenella ROP41 recombinant protein vaccine; and by immunizing chicks through the vaccine, the infection of the Eimeria tenella can be effectively resisted, the survival rate of the chicks can be increased, the relative weight gain rate of the chicks can be increased, and the discharge rate of oocysts of the Eimeria tenella can be decreased, so that the vaccine is a novel recombinant protein vaccine with a good immune protection effect and has the potential as a vaccine researched and developed for preventing a chicken Eimeria tenella disease.

The invention discloses a porcine circovirus type 2 Cap gene modified recombinant antigen and an application thereof, and belongs to the field of veterinary vaccine researches. The modified recombinant antigen is obtained by modification and construction of a Cap gene after a nuclear localization sequence of a porcine circovirus type 2 LG strain widely prevalent in swinery in China at present is deleted. A vaccine prepared from the modified recombinant antigen can stimulate organisms to produce high-level neutralizing antibodies, is superior to commercially available inactivated influenza virus vaccines and separate Cap protein immunization groups after nuclear localization sequences are deleted, has a highest antibody level, is safe and harmless to immunized animals after inoculation, has the advantages of low price, safety, stability, high yield and easiness in preservation and application, and has great significance to related diseases caused by porcine circovirus type 2 infection in China, so that the porcine circovirus type 2 Cap gene modified recombinant antigen has a good application prospect.

The invention relates to a method for producing CCR5 autovaccine, wherein based on the structure of CCR5, it uses linker whose amino acid sequence is GGGGS to connect external four sections of CCR5 cell, to simulate the external structure as rCCR5; inserts T cell assist epitope PADRE into N end of rCCR5; artificially synthesizes PADRE- rCCR5 gene; the target gene is colon into represent carrier of pBV-220 corn; transfers bacillus coli, to represent effect in it; washes with inclusion body; chromatography in gel post; obtaining purified protein; the target protein PADRE-rCCR5 is used as CCR5 polypeptide vaccine antigen, while immunity animal represents that the PADRE-rCCR5 can induce body to generate the antibody with high CCR5 specificity; and the invention uses flow cell check immunity antiserum to test U937 cell of CCR5, which proves that the average combine rate can reach 75. 8%. The invention supports CCR5 autovaccine construction and AIDS vaccine research.