37 results about "Tetanus Toxins" patented technology

The invention discloses a recombinant tetanus toxin protein rTT as well as a preparation method and application thereof, the recombinant protein sequentially contains a His tag, a modified exogenous polypeptide and two tetanus toxin fragments connected in series through flexible polypeptides from the N end to the C end, the two tetanus toxin fragments connected in series form two intramolecular disulfide bonds in a molecule through double mutation, and the tetanus toxin fragments are connected in series through flexible polypeptides. Coding nucleotide of the recombinant protein is expressed by escherichia coli BL21 (DE3), induced at low temperature and purified, and target protein with the purity larger than or equal to 95% can be obtained. A subunit vaccine prepared from the protein and a two-way oil adjuvant ISA201 enables mice to achieve 100% protection under 100 LD50 challenge dose, the protection effect is better than that of truncated tetanus toxin heavy chain fragment TTc protein, and the recombinant protein can be used for tetanus prevention vaccines and serological diagnostic reagents.

Disclosed in the present invention is a pharmaceutical combination comprising an adsorbed tetanus vaccine and an antibody or an antigen-binding fragment thereof against tetanus toxin. The pharmaceutical combination is used for preventing and / or treating tetanus.

The present invention relates to a nucleic acid molecule for modulating neuromuscular transmission, said nucleic acid molecule comprising a sequence encoding one or more motor endplate receptors or fragments thereof. The present invention further relates to a fusion protein, preferably comprising one or more features of one or more of the foregoing Claims, said fusion protein comprising a motor endplate receptor and / or fragments thereof, wherein said motor endplate receptor is an acetylcholine receptor (AChR), a muscle-specific tyrosine kinase (MuSK) receptor, and / or a glutamate receptor, and / or fragments thereof, and / or a neurotoxin, wherein said neurotoxin is botulinum toxin (BoNT) and / or tetanus toxin.

The invention discloses a preparation method of a proteoglycan protein conjugate carrying a specified connection site. The method comprises the following steps: carrying out recombinant expression on a recombinant tetanus toxin heavy chain C fragment rTTHc containing a Sortase A 7M ligase recognition sequence, introducing O, O '-(ethane-1, 2-diyl) bis (hydroxylamine) as a nucleophilic reagent into the C tail end of the rTTHc under the action of ligase Sortase A 7M, and carrying out specific condensation reaction on the C tail end of the rTTHc as a unique chemical reaction site and polysaccharide carrying an aldehyde group functional group to obtain the tetanus toxin heavy chain C fragment rTTHc. Therefore, polysaccharide protein conjugates with definite polysaccharide and protein connection sites are produced. According to the invention, the problem that the key antigen epitope of the carrier protein and the heterogeneity of the product are possibly damaged by the traditional random modification strategy is solved, the antigen epitope of the carrier protein is prevented from being damaged to a great extent, and a reliable technical scheme is provided for preparing the polysaccharide conjugate vaccine with high quality and high immunogenicity.

The invention provides an anti-tetanus antibody as well as preparation and application thereof, and relates to the field of antibodies. The amino acid sequence of the tetanus-resistant antibody heavy chain CDR is as shown in SEQ ID NO: 1-3; the amino acid sequence of the light chain CDR of the antibody is as shown in SEQ ID NO: 4-6. The antibody disclosed by the invention is high in activity and affinity, and has the application prospect of preparing a medicine for preventing and / or treating tetanus or a product for detecting the tetanus toxin level with a non-diagnostic purpose and the like.

Provided herein are antigenic peptides comprising the SARS-COV-2 spike protein receptor binding domain (CRBD) polypeptide or portions thereof, linked to a non-catalytic, non-toxic tetanus toxin variant (i.e., a modified tetanus toxin or “MTT”) and vaccine compositions comprising the same. In addition, provided herein are methods for making and using CRBD-MTT fusion proteins as immunogenic agents.

The present disclosure provides an anti-tetanus toxin neutralizing antibody as a new generation of tetanus passive immunization preparation. The antibody can be fully human, which can specifically bind to full-length tetanus toxin and its AB fragment, has high affinity, high specificity, small toxic side effects, avoids the problems of large adverse reactions and short half-life of animal-derived monoclonal antibodies, and is prepared in large quantities by mammalian cells, and has clear and stable components.

Disclosed in the present invention is a formulation containing an anti-tetanus toxin antibody, wherein the formulation comprises the anti-tetanus toxin antibody at a concentration of 1-100 mg / mL, and has a pH of 5.0-7.0; and optionally, the formulation further comprises one or more of a surfactant, a pH regulator, a protein protective agent, and an osmotic pressure regulator. The formulation provided by the present invention can effectively maintain the stability of the antibody, so that the antibody is not liable to aggregate, thereby preventing oxidation, denaturation and degradation. In addition, the formulation also maintains the high titer of the antibody against tetanus toxin, and thus has the high biological activity required for preventing or treating tetanus.

Provided are a bispecific antibody and a monoclonal antibody for a tetanus toxin, and uses of the antibodies. The bispecific antibody contains a first antigen-binding fragment and a second antigen-binding fragment which bind to different epitopes of the tetanus toxin, and has activity of neutralizing the tetanus toxin. The monoclonal antibody binds to the tetanus toxin, and has activity of neutralizing the tetanus toxin.

The application discloses a preparation containing an anti-tetanus toxin antibody, the preparation containing the anti-tetanus toxin antibody at a concentration of 1-100 mg / mL and having a pH of 5.0-7.0; and optionally, the preparation further contains one or more of a surfactant, a pH regulator, a protein protective agent and an osmotic pressure regulator. The preparation provided by the application can effectively maintain the stability of the antibody, so that the antibody is not prone to aggregation, oxidation, denaturation and degradation; in addition, the antibody has a high titer of anti-tetanus toxin and high bioactivity required for preventing or treating tetanus.

The invention discloses a nano antibody T92-6 of an anti-tetanus toxin Hc structural domain and application of the nano antibody T92-6, and belongs to the field of immunotherapy biotechnology pharmacy. The nano antibody or the antigen binding fragment thereof comprises three complementarity determining regions CDR1, CDR2 and CDR3, the amino acid sequence of the CDR1 is SEQ ID NO: 6, the amino acid sequence of the CDR2 is SEQ ID NO: 7, the amino acid sequence of the CDR3 is SEQ ID NO: 8, and the sequences of the complementarity determining regions are defined according to an IMGT numbering system. The antibody is only combined with THc protein, and poisoning caused by a lethal dose of tetanus toxin (10 * LD50) can be effectively blocked by 5 micrograms of the antibody; after being humanized, the antibody still has better affinity and neutralizing activity, and the antibody with the same dose can still effectively block poisoning caused by tetanus toxin (10 * LD50) with a lethal dose.

The application discloses a non-toxic genetically engineered toxoid molecule antigen vaccine based on a protective antigen functional domain structure and a preparation method and application thereof. It is determined that botulinum toxin and tetanus toxin both have two key protective antigen molecules, i.e. a receptor binding region Hc and a light chain-transmembrane region L-HN. Based on the important role of the functional domain in immunoprotective efficacy, a non-toxic genetically engineered toxoid or chimeric toxoid multivalent vaccine molecule is designed and prepared by using a biosynthesis technology. The vaccine has two important protective antigens of biological toxins, can produce strong complete protection at a low dose and a small number of immunization times, and can be used as a high-efficiency genetically engineered toxoid vaccine to replace formaldehyde inactivated toxoids for biological toxin prevention. The vaccine has important protective antigens of two different biological toxins, can produce protective efficacy against a plurality of different toxin biological agents, and can be used as a broad-spectrum multivalent vaccine for biological toxin prevention.

The invention provides a nanoparticle protein rePO-TTFC (at) LS as well as a preparation method and application thereof. The protein is composed of rePO (at) LS and a tetanus toxin C fragment TTFC. The recombinant protein SpyCatcher-rePO at LS disclosed by the invention is subjected to induced expression in escherichia coli in a soluble form. According to the present invention, the recombinant protein SpyCatcher-rePO (at) LS can be subjected to covalent binding with the recombinant protein reTTFC-SpyTag through the isopeptide bond so as to form the rePO-TTFC (at) LS duplex nanoparticle vaccine molecule, and the binding condition is simple; the nanoparticle protein can induce an animal body to generate high-level immune response and play an immune protection role, the response speed and protection efficiency of an antibody induced by the nanoparticle protein are obviously superior to those of a monomer rePO and a monomer reTTFC fusion protein, and the nanoparticle protein can be used for preventing and treating pseudomonas aeruginosa and tetanus at the same time.

The present invention relates to the field of biomedical technologies, and more specifically to a modified mRNA vaccine against tetanus and its preparation method. A modified mRNA vaccine against tetanus, characterized in that: the vaccine comprises: (1) mRNA for expressing a tetanus toxin immunogen, the immunogen being one of an Hc protein, an Hc-TRX protein, and an Hn+Hc protein; (2) a vaccine carrier, the vaccine carrier being a liposome nanoparticle that can be utilized by the mRNA. The main advantages of the present invention are: (1) for the first time, the C-terminal domain of the heavy chain of tetanus toxin (Hc), the fusion protein of the C-terminal domain of the heavy chain and thioredoxin (TRX-Hc), and the complete heavy chain of tetanus toxin (Hn+Hc) are selected as antigens to develop a modified mRNA vaccine; (2) the three developed tetanus toxin mRNA vaccines can all effectively induce specific antibody responses.

The invention discloses a nano antibody TL-25 of an anti-tetanus toxin L-HN fragment and application of the nano antibody TL-25, and belongs to the field of immunotherapy biotechnology pharmacy. The nano antibody or the antigen binding fragment thereof comprises three complementarity determining regions CDR1, CDR2 and CDR3, the amino acid sequence of the CDR1 is SEQ ID NO: 6, the amino acid sequence of the CDR2 is SEQ ID NO: 7, the amino acid sequence of the CDR3 is SEQ ID NO: 8, and the sequences of the complementarity determining regions are defined according to an IMGT numbering system. The antibody is only combined with TL-HN protein, and 0.3125 [mu] g of the antibody can effectively block poisoning caused by tetanus toxin (10 * LD50) in a lethal dose; after being humanized, the antibody still has better affinity and neutralizing activity, and the antibody with the same dose can still effectively block poisoning caused by tetanus toxin (10 * LD50) with a lethal dose.

Provided are an anti-tetanus toxin neutralizing antibody and the use thereof as a new-generation tetanus passive immunological preparation. The antibody can be a fully human antibody, has the characteristics of a high affinity, a high specificity, and few toxic side effects, and avoids the problems of strong adverse reactions and a short half-life of animal-derived monoclonal antibodies. Moreover, the antibody can be prepared on a large scale using mammalian cells, and has a clear and stable composition. On the basis of clinical requirements, the exploration and development of an anti-tetanus toxin neutralizing fully-human antibody as a new-generation tetanus passive immunological preparation has great medical value.

This invention provides a means to inactivate the inherent toxicity of tetanus toxin and produce a safe and effective vaccine. [Solution] This specification provides engineered non-catalytic, non-toxic tetanus toxin variants and methods for using such engineered tetanus toxin variants as low-dose prophylactic vaccines that are more effective than non-toxic and their respective chemically inactivated toxoids. In addition, this specification provides conjugate vaccine carriers containing engineered tetanus toxin variants and methods for using such conjugate vaccines to induce a T-cell-dependent immunomemory response that can target a broad range of microbial pathogens as a single vaccine.

The invention discloses a preparation and test method of an immunosensor for tetanus toxin detection. The method comprises the following steps: respectively preparing a tetrachloroauric acid solution and a hexachloroplatinic acid solution, dropwise adding the tetrachloroauric acid solution and the hexachloroplatinic acid solution to the red copper sheet, and drying under an ultraviolet lamp to form a gold-platinum nanoparticle catalyst layer; placing the red copper sheet in a chamber of electron-assisted hot filament chemical vapor deposition system equipment for growing a gold platinum-vertical graphene electrode; dropwise adding a tetanus antibody solution and a bovine serum albumin solution into the gold platinum-vertical graphene electrode to obtain a gold platinum-vertical graphene electrode incubated with bovine serum albumin and a tetanus antibody; a tetanus toxin solution is taken and dropwise added to the gold platinum-vertical graphene electrode incubated with bovine serum albumin and tetanus antibodies, and the immunosensor for tetanus toxin detection is obtained. The method inhibits non-specific adsorption, stabilizes a base line, and supports tetanus toxin low-concentration quantification and subsequent application linkage.

The invention discloses a method and a product for detecting the binding capacity of clostridium neurotoxin and a receptor. The research discovers that the synaptic binding protein Syt1, a Syt1 mutant or Syt2 can be used for promoting glycosylation modification and membrane localization of the synaptic vesicle protein SV2, so that the binding capacity of the synaptic vesicle protein SV2 and botulinum neurotoxin or tetanus toxin is remarkably enhanced. The synaptic binding protein Syt1, Syt1 mutant or Syt2 is prepared into a product for detecting the clostridium neurotoxin, so that the synaptic vesicle protein SV2 can retain natural conformation and post-translational modification sites, and the physiological expression state is effectively simulated, thereby realizing accurate analysis of the binding capacity of the clostridium neurotoxin and the synaptic vesicle protein SV2.

The invention provides a pharmaceutical composition for preventing and / or treating tetanus as well as a preparation method and application of the pharmaceutical composition. The pharmaceutical composition comprises a recombinant tetanus toxin C fragment as a unique antigen component, an adjuvant and a solvent, wherein the adjuvant is selected from one or more of an aluminum adjuvant, a Toll-like receptor stimulant and saponin, preferably the aluminum adjuvant, and / or the solvent is selected from one or more of a phosphate buffer solution, a citrate buffer solution, a histidine salt buffer solution and a sodium chloride solution, preferably the phosphate buffer solution and / or the sodium chloride solution. The adopted recombinant tetanus natural C fragment (Hc) is non-toxic, formaldehyde inactivation is not needed, meanwhile, the possibility of toxicity reversion is avoided, and the pharmaceutical composition can induce mammals to generate high immune response against tetanus and has good immune persistence.

The invention discloses a method for extracting tetanus toxin through salting-out separation and application. The method comprises the following steps: carrying out first-stage salting-out on a clostridium tetani fermentation liquor by using ammonium sulfate, carrying out first-stage membrane filtration, collecting a supernatant A, carrying out second-stage salting-out on the supernatant A by using ammonium sulfate, carrying out second-stage membrane filtration, and collecting a precipitate B; wherein a membrane for the first-stage membrane filtration comprises a composite filter membrane with the pore size of 3.0-5.0 mu m and a composite filter membrane with the pore size of 0.3-0.5 mu m, a membrane for the second-stage membrane filtration comprises a filter membrane with the pore size of 0.6-1.0 mu m, and the precipitate B comprises tetanus toxin. The method can be used for treating a large-volume solution, remarkably shortening the process time, reducing the labor cost, realizing full-flow channelization operation, effectively preventing microbial pollution, facilitating linear amplification of the process, remarkably improving the early-stage purification effect of protein, removing a large amount of impurities and relieving the burden of subsequent purification steps.

The invention discloses an anti-tetanus toxin bispecific neutralizing antibody as well as a related biological material and application thereof, and belongs to the field of immunotherapy biotechnology pharmacy. The bispecific antibody or the antigen binding fragment of the bispecific antibody comprises two humanized nano antibodies, namely T92-6-h2 and TL-25-h1, and amino acid sequences of three complementary determining regions CDR1, CDR2 and CDR3 of the T92-6-h2 are SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8 in sequence; the amino acid sequences of three complementary determining regions CDR1, CDR2 and CDR3 of the TL-25-h1 are SEQ ID NO: 14, ID NO: 15 and SEQ ID NO: 16 in sequence; the sequence of the complementarity determining region is defined according to an IMGT numbering system. The antibody can be combined with an L-HN functional fragment and an Hc structural domain at the same time, and can effectively block poisoning caused by lethal dose tetanus toxin.

Provided is an application of an anti-tetanus toxin antibody in preventing or treating tetanus. The anti-tetanus toxin antibody is applied in a specific administration regimen, and can achieve the technical effects of being quicker to take effect and having a longer protection time than existing passive immunization agents, and can therefore effectively prevent or treat tetanus.

The present disclosure discloses a method for purifying tetanus toxin or variants thereof. The method for purifying tetanus toxin or variants thereof uses metal chelate affinity chromatography, and the tetanus toxin does not contain an affinity tag. The present disclosure uses biocompatible agarose-based nickel affinity chromatography and anion exchange chromatography as separation subjects, which can quickly and effectively remove impure proteins and enrich target proteins; it improves the separation and purification efficiency of proteins and greatly reduces the loss of proteins.

The invention discloses a preparation method of tetanus full-length toxoid molecules inactivated by a genetic engineering technology and application of the tetanus full-length toxoid molecules as vaccine antigens, and belongs to the field of biological pharmacy. The technical problem to be solved by the invention is how to obtain a novel tetanus full-molecular vaccine with enhanced immune effect and improved protective efficacy. The preparation method of the tetanus full-length toxoid molecule disclosed by the invention comprises the following steps: introducing an expression vector of a coding gene containing ddmTT protein into a microorganism to express and purify the coding gene, so as to obtain the tetanus full-length toxoid molecule, the ddmTT protein is the ddmTT protein as shown in SEQ ID No. 6 (sequence identifier number 6). The tetanus full-length toxoid molecule (ddmTT) prepared by the invention has the advantages of no toxicity, high safety, low production cost, strong immunogenicity, high protective efficacy and quick response, and can be used as a novel vaccine for preventing tetanus toxin poisoning, a novel immunogen and a novel carrier molecule of a coupling conjugate vaccine.

The application discloses a preparation and testing method of an immunosensor for detecting tetanus toxin. The method comprises the following steps: respectively preparing tetrachloroauric acid solution and hexachloroplatinic acid solution, and dropping the two solutions on a red copper sheet; after baking under an ultraviolet lamp, a gold platinum nanoparticle catalyst layer is formed; the red copper sheet is placed in a chamber of an electron-assisted hot-wire chemical vapor deposition system device, so as to grow a gold platinum-vertical graphene electrode; a tetanus antibody solution and a bovine serum albumin solution are dropped on the gold platinum-vertical graphene electrode, so that a gold platinum-vertical graphene electrode incubated with bovine serum albumin and tetanus antibody is obtained; a tetanus toxin solution is dropped on the gold platinum-vertical graphene electrode incubated with bovine serum albumin and tetanus antibody, so that an immunosensor for detecting tetanus toxin is obtained. The application can inhibit non-specific adsorption, stabilize a baseline, support low-concentration quantification of tetanus toxin, and connect subsequent applications.

The invention discloses a recombinant Marek's virus vector vaccine strain for expressing H9N2 subtype avian influenza virus HA protein and application of the recombinant Marek's virus vector vaccine strain, and belongs to the technical field of veterinary biological products. The amino acid sequence of the HA protein expressed by the recombinant Marek's virus vector vaccine strain is as shown in SEQ ID NO. 1. The amino acid sequence of the HA protein is obtained by modifying the 84th amino acid of the amino acid sequence shown as SEQ ID NO.2 into tryptophan from serine, eliminating the 82nd glycosylation site of the HA protein, replacing the signal peptide of the HA gene with the Ig lambda signal peptide of chicken, and adding GGGGS linker and one tetanus toxin T cell epitope to the carboxyl terminal of the HA protein. The vaccine strain is good in in-vitro proliferation characteristic, stable in genome heredity and capable of stably expressing HA protein, no side reaction or pathogenic effect exists after SPF chicken is immunized, 100% protection against H9N2 strain attack can be provided 21 days after immunization, and the problem that the immune protection blank period of other vaccines such as H9N2 inactivated vaccines is long is solved.