29 results about "Neutralizing epitope" patented technology

The invention relates to the technical field of biological medicine, in particular to a monoclonal antibody of bovine coronavirus S2 protein, an epitope of the monoclonal antibody and application of the epitope. The invention provides a hybridoma cell strain (preservation number: CCTCC (China Center For Type Culture Collection) NO: C202510). The cell strain can secrete a monoclonal antibody which is targeted and combined with bovine coronavirus S2 protein; the amino acid sequence of the epitope identified by the monoclonal antibody is shown as SEQ ID NO: 1. Experiments prove that the monoclonal antibody has neutralizing activity, and the targeting epitope of the monoclonal antibody is a key neutralizing epitope. The invention provides a new strategy and tool for antibody drug and vaccine research and development as well as diagnosis and detection of the bovine coronavirus.

The invention discloses a reconstructed respiratory syncytial virus F protein trimer, relates to the technical field of genetic engineering vaccines, and designs an RSV F protein trimer, a monomer of the trimer does not contain a p27 sequence, and a furin enzyme digestion peptide fragment aa110-aa136 is deleted; the F2 structural domain and the F1 structural domain are connected through a GS connexon; the invention also designs point mutation combinations, and the point mutation combinations can form an inter-chain disulfide bond, so that covalent connection of the trimer is realized, and the problem of depolymerization of the trimer is fundamentally solved; the trimer is stabilized in a pre-F state, a post-F sample structure does not exist, and an ELISA result shows that a neutralizing epitope is completely reserved; the trimer antigen structure is challenged by adopting a thermal acceleration experiment method, and a result shows that the purity of the trimer antigen reaches 95% or above after the trimer antigen is placed at 37 DEG C for 28 days, and the trimer antigen is very good in stability; when the trimer is used for preparing the vaccine, the standard required by laws and regulations can be met without freeze-drying.

The invention discloses an RSV pre-F tripolymer protein combination and application thereof in preparation of bivalent RSV vaccines, and relates to the technical field of human vaccines. The vaccine is prepared by mixing an RSV-A subtype pre-F tripolymer protein and an RSV-B subtype pre-F tripolymer protein in equal mass, and matching with an aluminum hydroxide adjuvant with the concentration of 0.35 mg / 0.5 mL. A p27 fragment and a C-terminal esterification region of each of the two F proteins are deleted, the two F proteins are connected with F2 / F1 through GSGSGS, inter-chain disulfide bonds are introduced to S146C / N460C and A149C / Y458C sites respectively, intra-chain disulfide bonds are introduced to S155C / S290C sites respectively, and stable pre-F trimers are spontaneously assembled after secretion expression of CHO cells; the vaccine does not need to be freeze-dried, still keeps high purity and neutralizing epitope integrity after being stored at 37 DEG C for 28 days, can obviously induce neutralizing antibody and T cell immunity, and can be used for preventing various RSV-A / B infections.

The invention discloses a multi-epitope fusion protein based on porcine rotavirus VP7 and VP8 double targets as well as a preparation method and application of the multi-epitope fusion protein. According to the invention, salmonella typhimurium flagellin FliCS.T is taken as a skeleton, dominant neutralizing epitopes of VP7 and VP8 are accurately integrated, and the multi-epitope fusion antigen FliCS.T-VP7 + VP8 is constructed. The fusion protein has two immunological advantages: an FliCS.T skeleton has TLR5 adjuvant activity, so that innate immunity and adaptive immunity of a host can be efficiently activated; key neutralizing epitopes of porcine rotaviruses VP7 and VP8 are synchronously presented through a multi-epitope strategy, and high-titer specific neutralizing antibody and potent cellular immune response aiming at two structural proteins can be induced in vivo. The invention provides the multi-epitope fusion antigen of the porcine rotavirus, which integrates high immunogenicity and broad-spectrum neutralization protection, and a feasible technical scheme is provided for prevention and control of the porcine rotavirus.

The invention discloses a porcine rotavirus VP8 protein dominant B cell neutralizing epitope peptide, a multi-epitope fusion protein and application of the multi-epitope fusion protein. The FliCS.T-VP8 multi-epitope fusion protein prepared by the invention is a multi-epitope fusion protein which takes salmonella typhimurium flagellin FliCS.T as a molecular skeleton and integrates the dominant B cell neutralizing epitope of the VP8 protein of the porcine rotavirus. According to the design, immunodominance epitope screening, multi-epitope fusion protein construction and genetic engineering technologies are fused, and the immunogenicity and immune protection efficacy of the fusion protein are remarkably enhanced. The FliCS.T-VP8 multi-epitope fusion protein not only can effectively induce high-level VP8 protein specific antibody response and cellular immune response, but also shows good porcine rotavirus neutralizing activity. In addition, the fusion protein has relatively high safety and stability, and has the potential of being used as an efficient subunit vaccine for preventing and controlling porcine rotavirus infection in the pig industry.

This invention provides a broad-spectrum mRNA vaccine for the prevention of feline infectious peritonitis (FIP) and its preparation method. The vaccine comprises mRNA molecule 1 and mRNA molecule 2. mRNA molecule 1 contains an LS-FIPV-I MEV-N-M antigen protein coding sequence composed of a self-assembled LS nanoparticle fusion sequence based on a tandem neutralizing epitope of the FIPV type I S protein and a T-cell epitope, as well as antigen-coding sequences of the N and M proteins. mRNA molecule 2 contains an antigen protein coding sequence of an LS nanoparticle structure self-assembled from the FIPV type II S protein receptor domain RBD. This invention, through the combination of two mRNA molecules, can achieve broad-spectrum protection against both FIPV types I and II while mitigating the risk of adverse drug reaction (ADE).

The invention discloses a high-stability and informatization-adaptive chemiluminescence mixed indoor quality control product for infectious disease projects and a preparation method thereof. The method is characterized in that inactivated healthy human serum is used as a non-infectious optimized matrix, a single kit covers all infectious disease A and B immunodetection items specified by the National Health Committee, and three concentration gradients of negative, weak positive and strong positive are set. Through cooperation of a recombinant non-neutralizing epitope antigen, a high-specificity monoclonal antibody and a double-blocking system, interference-free coexistence of multiple items is realized, and the cross reaction rate is less than or equal to 1.0%; a four-dimensional synergistic compound long-acting stable system is adopted, and it is ensured that the activity retention rate is larger than or equal to 95.0% after 24 months of storage at 2-8 DEG C. The product is packaged in a special direct on-machine test tube, is matched with a fixed identity code of a pre-associated laboratory information system, supports two-way communication with an automatic instrument, and realizes no manual intervention in the whole process of quality control. The preparation process is standardized, is suitable for clinical laboratories of all levels of medical institutions, and promotes import and substitution of high-end quality control products.

The invention belongs to the technical field of preparation of veterinary vaccines, and particularly relates to a Senecavirus type A nanoparticle antigen and application thereof in preparation of a nanoparticle vaccine. According to the invention, a general Th cell epitope is fused with a highly conservative B cell neutralizing epitope VP2150-160aa of a three-copy Senecavirus A capsid protein, and the fused recombinant protein is displayed on the surface of a nanoparticle, so that the Senecavirus A nanoparticle antigen is finally constructed. The Senecavirus A nanoparticle vaccine is prepared from the Senecavirus A nanoparticle antigen, and the Senecavirus A nanoparticle vaccine can display more antigens at a time, so that the immunogenicity of the vaccine can be improved, and the immune effect of the vaccine can be enhanced.

The invention discloses a preparation method and application of a monoclonal antibody based on recombinant goose circovirus Cap protein, and belongs to the technical field of biology. The monoclonal antibody prepared based on the recombinant goose circovirus Cap protein has excellent specificity and affinity, has excellent reactivity with the goose circovirus, and also has high specificity and sensitivity when being applied to preparation of diagnostic reagents. Besides, the prepared monoclonal antibody is applied to non-therapeutic purpose neutralization virus activity and passive immune activity, and the monoclonal body can block the infection mechanism by combining virus key protein (Cap protein); the antibody targets the neutralizing epitope of the virus capsid protein and inhibits virus replication; instant immune protection can be provided by directly injecting the monoclonal antibody, interaction with virus protein can be achieved, neutralizing epitopes can be accurately positioned, vaccine design can be guided, the monoclonal antibody can be used for quantitative detection of the antibody level after vaccine immunization, and the monoclonal antibody has important significance in virus diagnosis and illness state evaluation.

This invention provides a broad-spectrum non-neutralizing antibody against foot-and-mouth disease virus (FMD) and its application in the preparation of reagents that enhance the production of neutralizing antibodies, belonging to the field of antibody technology. The FMD virus broad-spectrum non-neutralizing antibody has the amino acid sequence of its heavy chain variable region as shown in SEQ ID NO:1 and the amino acid sequence of its light chain variable region as shown in SEQ ID NO:2. This invention utilizes single B-cell antibody technology to screen and obtain a porcine broad-spectrum non-neutralizing monoclonal antibody SP1C6, which can recognize a conformational epitope on VP2. The broad-spectrum non-neutralizing antibody SP1C6 increases the level of neutralizing antibodies induced by FMD virus in pigs by blocking the non-neutralizing epitopes of the FMD virus antigen.

The present disclosure provides antibodies that specifically bind to botulinum neurotoxins (e.g., BoNT / A, BoNT / B, BoNT / C, BoNT / D, BoNT / E, BoNT / F, BoNT / G, etc.) and the epitopes bound by those antibodies. The antibodies and derivatives thereof that specifically bind to the neutralizing epitopes provided herein can be used to neutralize botulinum neurotoxin and are therefore also useful in the treatment of botulism.

The invention discloses a recombinant expression vector as well as a construction method and application thereof. The invention also discloses a recombinant strain as well as a construction method and application thereof. The invention also discloses a genetic engineering live vector vaccine. According to the invention, probiotic bacillus subtilis is used as an antigen delivery carrier, and recombinant bacillus subtilis for recombinant integration tandem expression of porcine epidemic diarrhea virus antigen neutralizing epitopes and intestinal M cell and dendritic cell (DCs) targeting peptides is constructed. The recombinant live vector vaccine is orally taken and immunized by sows, and efficient intestinal mucosa immune protection is provided for piglets in the lactation period by enhancing the intestinal mammary axial function of the sows, so that a safe, simple and convenient immune way is provided for effectively preventing the occurrence of epidemic diarrhea of the piglets.

The application discloses a reconstructed respiratory syncytial virus F protein trimer, and relates to the technical field of genetic engineering vaccines.The application designs a RSV F protein trimer, the monomer of the trimer does not contain a p27 sequence, and a furin enzyme cleavage peptide segment aa110-aa136 is deleted; a GS linker is used for connecting between an F2 domain and an F1 domain; the application also designs point mutation combinations, the point mutation combinations can form inter-chain disulfide bonds, so that covalent connection of the trimer is realized, and the problem of trimer depolymerization is fundamentally solved; the trimer is stably kept in a pre-F state, and there is no post-F-like structure; an ELISA result shows that neutralizing epitopes are completely retained; a thermal acceleration experiment method is used to challenge the trimer antigen structure, and the result shows that the trimer antigen purity reaches more than 95% after being placed at 37 DEG C for 28 days, and the stability is very good; the trimer is used to prepare a vaccine, and the vaccine can meet the standard of a regulation requirement without freeze drying.

This invention discloses an RSV pre-F trimer protein combination and its application in the preparation of a bivalent RSV vaccine, relating to the field of human vaccine technology. The vaccine is prepared by mixing equal masses of RSV-A and RSV-B subtype pre-F trimer proteins and adding 0.35 mg / 0.5 mL of aluminum hydroxide adjuvant. Both F proteins have their p27 fragment and C-terminal esterification region deleted, and F2 / F1 are linked by GSGSGS. Interchain disulfide bonds are introduced at S146C / N460C and A149C / Y458C sites, and intrachain disulfide bonds are introduced at S155C / S290C sites. After secretion and expression by CHO cells, the pre-F trimer spontaneously assembles into a stable pre-F trimer. The vaccine does not require lyophilization and maintains high purity and the integrity of neutralizing epitopes after storage at 37°C for 28 days. It can significantly induce neutralizing antibodies and T-cell immunity and can be used to prevent various RSV-A / B infections.

The application belongs to the technical field of biotechnology, and particularly relates to a recombinant antigen of American trypanosomiasis and a preparation method and application thereof. The application provides a recombinant antigen of American trypanosomiasis, wherein a plurality of neutralizing epitopes and complete amino acid sequences of Trypanosoma cruzi antigens are connected according to a specific order to construct a recombinant antigen capable of covering all stages of the life cycle of Trypanosoma cruzi. The recombinant antigen is complete in Chagas antibody recognition site coverage, can recognize and combine all types of antibodies in the Chagas chronic and acute infection period, and is not easily interfered by other substances in serum, and is high in specificity.

The application discloses a multi-epitope fusion protein based on porcine rotavirus VP7 and VP8 double targets and a preparation method and application thereof. S.T As a skeleton, the advantages of precisely integrating VP7 and VP8 neutralizing epitopes are constructed into multi-epitope fusion antigen FliC S.T -VP7+VP8. The fusion protein has two immunological advantages: FliC S.T The skeleton has TLR5 adjuvant activity, can efficiently activate the innate immunity and adaptive immunity of a host, and can synchronously present key neutralizing epitopes of porcine rotavirus VP7 and VP8 through a multi-epitope strategy, so that high-titer specific neutralizing antibodies and strong cellular immune responses against the two structural proteins can be induced in the body. The application provides a multi-epitope fusion antigen of porcine rotavirus which integrates high immunogenicity and broad-spectrum neutralization protection, and provides a feasible technical scheme for the prevention and control of porcine rotavirus.

Provided herein are compositions including a binding reagent for a Zika virus (ZIKV) epitope, pharmaceutical compositions including such binding reagents, and methods of detecting ZIKV binding antibodies and ZIKV neutralizing antibodies in patients.

The invention provides a neutralizing epitope polypeptide of grouper nervous necrosis virus and application of the neutralizing epitope polypeptide in serological diagnosis test paper, and the amino acid sequence of the neutralizing epitope polypeptide is SEQ ID NO: 1 or SEQ ID NO: 2. On the other hand, the invention also provides a serological diagnosis test strip for the nervous necrosis virus disease of the grouper, wherein the NNV capsid protein neutralizing epitope polypeptide is used as a detection line. The antigen polypeptide used by the test strip is an NNV capsid protein neutralizing epitope specific fragment, the antigen recognition effect is improved by optimizing the mixing ratio, cross reaction with other pathogen positive serum of grouper is avoided, and the detection accuracy is ensured.

The application provides a method for identifying neutralizing antigen epitopes in bovine coronavirus spike protein, immunogens, compositions and uses, relates to the technical field of biology, and specifically constructs an efficient neutralizing epitope identification route: online prediction, nanoparticle multivalent display, animal immunization, combined evaluation of binding and neutralization, which can screen out epitopes with neutralizing activity from candidate linear epitopes in a relatively short period of time; the multivalent presentation of ferritin nanoparticles significantly improves the detectability of the immune response of weak immunogenic linear peptides; the obtained epitopes can directly serve the development of subunit vaccines, multi-epitope nanovaccines and in vitro diagnostic reagents.

The invention provides a broad-spectrum mRNA vaccine for preventing feline infectious peritonitis and a preparation method thereof, the vaccine comprises an mRNA molecule 1 and an mRNA molecule 2, the mRNA molecule 1 comprises a self-assembled LS nanoparticle fusion sequence based on a neutralizing epitope of series FIPV type I S protein and a T cell epitope, and the mRNA molecule 2 comprises a self-assembled LS nanoparticle fusion sequence based on series FIPV type I S protein. The LS-FIBV-I MEV-N-M antigen protein coding sequence is composed of the antigen coding sequences of the N protein and the M protein; the mRNA molecule 2 comprises an antigen protein coding sequence which is fused and self-assembled by an FIPV II type S protein receptor structural domain RBD and has an LS nano-particle structure. According to the invention, through the combination of double mRNA molecules, while the ADE risk is avoided, the broad-spectrum protection of the FIPV type I and the FIPV type II is realized.

The present invention discloses a monoclonal antibody that broadly binds to the HA protein of the multi-branched H5Nx subtype avian influenza virus and its application. The monoclonal antibody is named 1G10, belongs to the IgG1 subclass, comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises amino acid sequences as shown in SEQ ID NO.5-7. H CDR1, V H CDR2 and V H CDR3, the light chain variable region comprises amino acid sequences as shown in SEQ ID NO.8-10 L CDR1, V L CDR2 and V L The monoclonal antibody specifically recognized the HA protein in indirect immunofluorescence assays, protein immunoblotting, and neutralization tests. Furthermore, the monoclonal antibody was able to undergo specific hemagglutination inhibition with H5Nx subtype avian influenza viruses, including H5N1, H5N6, and H5N8, from different HA gene evolutionary branches, such as 2.3.2.1d, 2.3.4.4b, and 2.3.4.4d, indicating that the antigenic epitope recognized by the 1G10 monoclonal antibody is a broad-spectrum neutralizing epitope for H5 subtype avian influenza viruses. Chicken embryo immune escape assays and sequence alignment revealed that the neutralizing epitope recognized by the 1G10 monoclonal antibody includes amino acid positions 170 and 231 of the HA protein, and that mutations significantly affect antigenicity.

The invention provides a method for identifying a neutralizing epitope of bovine coronavirus spike protein, an immunogen, a composition and application, and relates to the technical field of biology, the method specifically constructs an efficient neutralizing epitope identification route: on-line prediction, nanoparticle multivalent display, animal immunization, and combined evaluation of associativity and neutrality. Epitopes with neutralizing activity can be screened out from candidate linear epitopes in a short period; the multivalent presentation of the ferritin nanoparticles significantly improves the immune response detectability of the weak immunogenicity linear peptide fragment; the obtained epitope can be directly used for development of subunit vaccines, multi-epitope nano vaccines and in-vitro diagnostic reagents.

This invention provides a modified hemagglutinin (HA) protein for preparing a broad-spectrum vaccine against H9 avian influenza, a broad-spectrum vaccine based on the HA protein, a preparation method, and applications. The modified HA protein is obtained through the following strategy: consensus sequences are constructed as immunogenic backbones for the HA sequences of H9 avian influenza viruses at different epidemic periods; based on this, two types of functional mutations are directionally introduced: one type involves transplanting key epitope residues targeted by known broad-spectrum antibodies, and the other type involves neutralizing epitope residues predicted based on viral evolutionary patterns. The modified HA protein of this invention can maintain a highly similar overall fold and trimer interface to the wild type, can be efficiently expressed in eukaryotic systems, and can induce high levels of neutralizing antibodies with broad-spectrum cross-protective effects in target animals. It provides a core antigen candidate for the development of a new generation of H9 broad-spectrum vaccines and has broad application prospects.

The present disclosure provides antibodies that specifically bind to botulinum neurotoxins (e.g., BoNT / A, BoNT / B, BoNT / C, BoNT / D, BoNT / E, BoNT / F, BoNT / G, etc.) and the epitopes bound by those antibodies. The antibodies and derivatives thereof that specifically bind to the neutralizing epitopes provided herein can be used to neutralize botulinum neurotoxin and are therefore also useful in the treatment of botulism.

The invention relates to the technical field of biological medicines and vaccines, in particular to a multi-epitope fusion antigen, a protein, a vaccine and application thereof. The amino acid sequence of the multi-epitope fusion antigen is as shown in SEQ ID NO: 1. According to the present invention, by screening the conservative neutralizing epitope in the SARS S2 region and reasonably designing the multi-epitope fusion antigen, the problem of insufficient protection of the existing novel coronavirus vaccine on the variant is solved, the broad-spectrum protection of the various novel coronavirus variants is achieved, and the broad application prospect is provided.

The invention relates to the technical field of biological medicines and vaccines, in particular to a multi-epitope peptide based on novel coronavirus and application of the multi-epitope peptide. The multi-epitope peptide is formed by connecting neutralizing epitope peptides with amino acid sequences as shown in SEQ ID NO: 1-11 through flexible connecting peptides. According to the invention, by screening the conservative neutralizing epitope in the SARS-COV2 S2 region and reasonably designing the multi-epitope fusion protein, the problem that the existing novel coronavirus vaccine is insufficient in protection of variants is solved, the broad-spectrum protection of various novel coronavirus variants is realized, and the application prospect is wide.

The invention relates to the technical field of biology, in particular to an HTLV recombinant antigen and a preparation method and application thereof. From the N terminal to the C terminal, the HTLV recombinant antigen comprises a P24 holoprotein, a gp46 holoprotein, a gp46 neutralizing epitope aggregation region and a gp21 extracellular domain which are connected in sequence, and the C terminal is connected or not connected with a tag. According to the present invention, the P24 holoprotein, the gp46 holoprotein, the p46 neutralizing epitope aggregation region and the gp21 extracellular domain are sequentially connected so as to form the HTLV recombinant antigen with the specific structure, and the positive coincidence rate and the negative coincidence rate are high when the HTLV antibody in the sample is detected by using the recombinant antigen;

The invention provides an African swine fever virus-like particle and application thereof in preparation of an African swine fever virus vaccine, and relates to the technical field of biology. The capsid protein, the protein A, the protein B and the protein C provided by the invention have good immune effects and can be used in African swine fever virus vaccines. Wherein the protein C is a four-layer nested particle and maintains a three-dimensional conformation of neutralizing epitopes on the surface of the particle, so that the structural rigidity of the particle is enhanced, and a better immune effect is achieved.

The application discloses a preparation method of a PEDV COE subunit vaccine targeting M cells and application thereof. The application connects RGD peptide targeting M cells, a core neutralizing epitope COE region on a PEDV S protein and a fluorescent protein, expresses a recombinant protein, introduces a sulfur-containing amino acid into the protein to facilitate modification of the recombinant protein and connection of the protein to other substances in a covalent bond mode, and obtains a safe and effective mucosal subunit vaccine with green fluorescent labeling. Unlike common vaccines, the antigen of the vaccine can not only be directly taken by dendritic cells, but also be presented to dendritic cells through targeting M cells, the antigen presentation efficiency is significantly improved, the downstream immunity is more effectively started, and a series of immune responses of the body are induced, so that the mucosal immune system of the body produces a large number of PEDV specific antibodies to resist invasion of external pathogens, and the effect of specific immune protection is achieved.