45 results about "Tetanus Toxins" patented technology

The invention provides a chemically modified tetanus toxin C fragment as well as preparation and application thereof. The method for chemically modifying the tetanus toxin C fragment comprises the following steps: 1) reacting the tetanus toxin C fragment with a reducing agent to reduce a disulfide bond in the tetanus toxin C fragment; and 2) carrying out a reaction on the reduced tetanus toxin C fragment and an alkylation reagent, and carrying out modification. The recombinant tetanus toxin C fragment prepared by the method disclosed by the invention not only meets the quality standard required by pharmacopeia, but also has the advantages of stable conformation, high yield, easiness in amplification and low cost, and has a wide application prospect in the preparation of human vaccines, veterinary vaccines and antibody drugs and the development process of in-vitro diagnostic reagents.

The invention provides an application of an anti-tetanus toxin antibody in prevention or treatment of tetanus. The anti-tetanus toxin antibody is applied according to a specific administration scheme, and the technical effects that the anti-tetanus toxin antibody takes effect more quickly and the protection time is longer than that of an existing passive immune preparation can be achieved, so that tetanus can be effectively prevented or treated.

The present invention provides a pharmaceutical combination comprising a tetanus-adsorbing vaccine and an antibody against tetanus toxin or an antigen-binding fragment thereof. The pharmaceutical composition is used for preventing and / or treating tetanus. According to the present invention, the combination of the recombinant natural completely humanized monoclonal antibody and the tetanus adsorption vaccine proves that the detection time of the antibody induced by the tetanus adsorption vaccine is advanced, and the high titer level can last for a longer time;

The invention discloses a tetanus toxoid refining method which comprises the following steps: S1, centrifuging tetanus fermentation harvest liquid by adopting a butterfly centrifuge to remove thalli, and then performing ultrafiltration concentration to obtain a concentrated solution; s2, treating the concentrated solution obtained in the step S1 with a D390 ion exchange chromatographic column to obtain a chromatographic solution; s3, treating the chromatographic solution obtained in the step S2 by virtue of a Superdex 200 ion exchange chromatographic column, eluting by virtue of a sodium carbonate buffer solution, and collecting an elution peak, namely target protein, so as to obtain an eluent; and S4, performing formaldehyde detoxification treatment on the eluent obtained in the step S3, performing ultrafiltration to remove formaldehyde, and finally performing sterilization through a filter membrane to obtain the toxoid stock solution. In the refining process, the loss of the target protein is less, and the prepared toxoid is relatively excellent in immune effect and relatively high in purity.

The present invention relates to antibodies, and antigen binding fragments thereof, that bind to tetanus toxin. The invention also relates to nucleic acids that encode, and to cells that express such antibodies and antibody fragments. In addition, the invention relates to the use of the antibodies and antibody fragments in the diagnosis, prophylaxis and therapy of infection with Clostridium tetani or tetanus.

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.

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 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 present invention is directed to a composition, mammalian cell and vector, and a method for the production of, and method of treatment with, a recombinant protein vaccine in a mammalian cell line. The methods and compositions are particularly useful for generating the stable expression of a recombinant protein vaccine of interest. The invention is particularly useful for the production of vaccines to aid in protection against viral pathogens for vertebrates, in particular mammalians, especially humans. The mammalian cell for producing a protein of interest comprises: a plasmid encoded with a nucleotide sequence encoding one or more epitopes or subunits of the SARS-CoV-2 that are embedded in the nucleotide sequence encoding a detoxified recombinant tetanus toxin (DrTeNT).

The invention discloses an anti-tetanus toxin L-HN fragment nano antibody as well as a related biological material and application thereof, 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.

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.