30 results about "Botulinum toxin type" patented technology

The invention discloses a detection product and method for detecting botulinum toxin type A component protein. The detection product provided by the invention comprises an antibody capable of specifically recognizing the botulinum toxin type A component protein. The antibody disclosed by the invention has good affinity and stability, can be widely applied to detection of botulinum toxin component protein A, can be combined with a capillary immunoelectrophoresis method to establish a standard map for detecting botulinum toxin component antibodies, and provides a quality control means for commercialized and self-researched antibodies, so that the antibody has good application prospects. According to the method, more possibilities are provided for establishment of related botulinum toxin detection methods, and detection requirements of more scenes can be met.

The invention discloses a detection method for detecting botulinum toxin A HA-70 and a product. The detection product disclosed by the invention comprises an antibody capable of being combined with the botulinum toxin A HA-70. The antibody has good affinity and stability, can be widely applied to detection of botulinum toxin type A HA-70, can be combined with a capillary immunoelectrophoresis method to establish a standard map for detection of botulinum toxin component antibodies, and provides a quality control means for commercialization and self-research of antibodies, so that the antibody has good application prospects. According to the method, more possibilities are provided for establishment of related botulinum toxin detection methods, and detection requirements of more scenes can be met.

The invention discloses a kit, an antibody and a method for detecting A-type botulinum toxin hemagglutinin-70. The kit of the present invention comprises an antibody capable of targeting type A botulinum toxin hemagglutinin-70. The antibody disclosed by the invention has good affinity and stability, can be widely applied to detection of A-type botulinum toxin hemagglutinin-70, can be combined with a capillary immunoelectrophoresis method to establish a standard map for detecting botulinum toxin component antibodies, and provides a quality control means for commercialized and self-researched antibodies, so that the antibody has good application prospects. According to the method, more possibilities are provided for establishment of related botulinum toxin detection methods, and detection requirements of more scenes can be met.

The invention discloses a preparation method and application of a recombinant A-type botulinum toxin, and aims to industrially produce the recombinant A-type botulinum toxin by utilizing the preparation method of the recombinant A-type botulinum toxin provided by the invention. The method comprises the following steps: firstly, constructing genetically engineered bacteria capable of heterologously expressing a botulinum toxin A gene in host bacteria by virtue of a genetic engineering means, then accumulating recombinant botulinum toxin A in a strain by virtue of the genetically engineered bacteria through high-density fermentation, collecting fermented thalli, crushing and purifying to obtain the recombinant botulinum toxin A, and finally, preparing the recombinant botulinum toxin A by virtue of the recombinant botulinum toxin A. And adding an auxiliary poising agent and a protective agent to prepare a liquid agent or a freeze-drying agent to obtain the recombinant A-type botulinum toxin for injection. According to the technical scheme, the complete recombinant botulinum toxin A with the purity as high as 99.9% is obtained, the yield is as high as 7000000 U / L, and the complete recombinant botulinum toxin A has no obvious difference from natural botulinum toxin A in structure and function and can be applied to multiple fields such as medical treatment, medical beauty and military affairs.

The application provides a toxicity-enhanced neurotoxin recombinant protein and application thereof, and belongs to the technical field of biological medicine. The recombinant protein comprises a botulinum toxin type A receptor binding domain and at least one GM1 binding peptide inserted into the botulinum toxin type A receptor binding domain, and the recombinin protein can recognize and bind to ganglioside GM1. The short peptide sequence capable of binding to ganglioside GM1 is introduced into the botulinum toxin type A receptor binding domain to obtain the recombinant protein, the binding capacity of the recombinant protein to ganglioside GM1 is enhanced, the target recognition and binding capacity of the recombinant protein to the nerve cell membrane are improved, the overall affinity and endocytosis efficiency of the recombinant protein to the nerve cell are improved, and the neurotoxicity of the botulinum toxin is enhanced. The application not only improves the binding efficiency of BoNT / A in the in-vitro nerve cell model, but also shows a higher toxicity level in the functional verification, and shows a good clinical conversion prospect.

The invention belongs to the field of biology, and particularly relates to a botulinum toxin A light chain mutant and application. The BoNT / A LC mutant is obtained by performing directed evolution optimization on the BoNT / A LC by adopting a phage-assisted evolution method, and compared with a wild type BoNT / A LC, the activity of the mutant is remarkably improved, the defects of an existing botulinum toxin product and the problems of drug resistance and the like can be overcome, and the BoNT / A LC mutant can be applied to the fields of medical beauty and medical treatment and has a wide application prospect.

The application discloses a kit, an antibody and a method for detecting botulinum toxin type A hemagglutinin-70. The kit comprises an antibody capable of targeting botulinum toxin type A hemagglutinin-70. The antibody has good affinity and stability, can be widely applied to detection of botulinum toxin type A hemagglutinin-70, and can combine with a capillary immunoelectrophoresis method to establish a standard atlas for detection of botulinum toxin component antibodies, and provide a quality control means for commercialized and self-developed antibodies. Therefore, the application provides more possibilities for establishment of a botulinum toxin related detection method, and can meet detection requirements of more scenes.

The invention relates to a recombinant botulinum toxin A1 precursor polypeptide and a method for preparing botulinum toxin A1 by using the same.

The present invention provides a recombinant botulinum toxin type A light chain, a recombinant botulinum toxin, and compositions, uses and methods related thereto, which have increased efficacy and half-life, improved patient convenience and allow for tailored treatment for specific indications.

The invention relates to the technical field of biological medicines, in particular to application of a small-molecule inhibitor MYF-03-176 in preparation of a medicine for treating and / or preventing keloid and the medicine. The MYF-03-176 can remarkably promote fibroblast apoptosis of keloid tissue and inhibit cell proliferation, the volume of the keloid tissue can be remarkably reduced by locally injecting the MYF-03-176 into an animal, the MYF-03-176 is expected to become an effective drug choice for treating the keloid, and the combined treatment effect of the MYF-03-176 and the A-type botulinum toxin is further improved.

This invention discloses an anti-botulinum toxin type A neutralizing antibody A16 and its related biomaterials and applications. This invention belongs to the field of biotechnology, specifically relating to anti-botulinum toxin type A neutralizing antibody A16 and its related biomaterials and applications. The nanobody targeting botulinum toxin type A or the antigen-binding fragment containing the nanobody of this invention has three complementary determinants CDR1, CDR2, and CDR3; wherein the amino acid sequence of CDR1 is shown in SEQ ID No. 2, the amino acid sequence of CDR2 is shown in SEQ ID No. 3, and the amino acid sequence of CDR3 is shown in SEQ ID No. 4. The fusion protein A16-hFc obtained by fusing the nanobody A16 with the Fc fragment (hFc) of human immunoglobulin can effectively block botulinum toxin type A infection, resisting 20LD... 50 ED50 of botulinum toxin 50 The value is 1.1 μg.

The application belongs to the technical field of medicine, and relates to a botulinum toxin type A mutant and application thereof. A series of botulinum toxin type A mutants are obtained by using a trinucleotide primer containing NNK at a mutation site, adopting site-directed mutagenesis technology to establish a single-site saturated mutation library and a combined mutation library, and screening. Compared with wild-type botulinum toxin type A, the botulinum toxin type A mutant has improved affinity with a receptor SV2C. The obtained full-length mutant toxin has improved activity of cutting a substrate SNAP-25 and muscle paralysis ability, reduced diffusion, significantly reduced lethal dose to mice, and improved safety, and can be applied to preparation of full-length BoNT / A mutant drugs for treating related diseases that can be treated by wild-type toxin and medical cosmetic related products. Meanwhile, the botulinum toxin type A mutant can be used as a drug carrier for drug delivery to the nervous system or an SV2 receptor.

The invention discloses a botulinum toxin type A heavy chain TD protein and application thereof. The amino acid sequence of the TD protein is as shown in SEQ ID No. 1, and the nucleotide sequence of the coding gene of the TD protein is as shown in SEQ ID No. 2. The botulinum toxin type A heavy chain TD protein is applied to preparation of anti-depression drugs, and is especially used for protecting neuronal functions and improving depression-like behaviors. The A-type botulinum toxin heavy chain TD protein effectively avoids the toxic effect of traditional A-type botulinum toxin, only effective fragments playing an anti-depression role in the A-type botulinum toxin heavy chain are extracted, and the effective anti-depression role is played by preparing small-molecule oligopeptide protein.

The present invention relates to use of botulinum toxin type E for preventing or treating neuropathic pain, and specifically, to a pharmaceutical composition comprising botulinum toxin type E for preventing or treating neuropathic pain, and a surgical operation analgesic comprising botulinum toxin type E as an active ingredient. The botulinum toxin type E of the present invention has an analgesic effect by significantly inhibiting neuropathic pain, and, after a single administration, the botulinum toxin type E of the present invention can alleviate side effects and provide an analgesic effect superior to that of Gabapentin, which is a conventional analgesic used for treating neuropathic pain, in mechanical allodynia caused by mandibular alveolar nerve injury. In addition, the botulinum toxin type E of the present invention has a short duration of analgesic action in the body after administration so as to enable faster recovery and rehabilitation than after administration of botulinum toxin type A, and thus can be effectively used for treatment of neuropathic pain.

The present invention relates to the technical field of a method for preparing a recombinant botulinum toxin type A, and particularly relates to a botulinum toxin type A obtained by means of gene recombination technology, the amino acid sequence of which is identical to the sequence of botulinum toxin type A prepared by means of extraction and purification from a wild Clostridium botulinum strain. Provided is a single-chain protein of botulinum toxin type A, sequentially comprising: a light chain of botulinum toxin type A or a modified light chain of botulinum toxin type A, a linking region, and a heavy chain of botulinum toxin type A. The single-chain protein is subjected to fermentation, crude purification, enzyme digestion, and fine purification to obtain high-quality recombinant botulinum toxin type A. The present invention can solve the existing problems of complex process, long period, high safety risk and the like in the preparation of botulinum toxin type A by using wild Clostridium botulinum strains, and also solve the existing problem that the amino acid sequence of botulinum toxin type A prepared by means of gene recombination technology cannot be identical to that of botulinum toxin type A prepared by wild Clostridium botulinum strains.

The present invention provides an application of botulinum toxin type A in the preparation of a drug for reducing local muscle damage to limbs after bites by Agkistrodon acutus, and relates to the technical field of snake venom bite drugs. This application confirms the application effect of botulinum toxin type A in the treatment of local muscle damage to limbs after bites by Agkistrodon acutus through research, and on this basis explores the research and development and clinical transformation of botulinum toxin type A drugs. Specifically, this application is obtained by applying a botulinum toxin type A preparation to the biting muscle tissue of an animal model of Agkistrodon acutus bite for four weeks. Compared with the animal model of Agkistrodon acutus bite, the botulinum toxin type A can effectively reduce the circumference of the animal limbs and the level of muscle damage markers in the serum after treatment, and in the analysis of the degree of pathological damage, the degree of apoptosis of muscle tissue cells, and the degree of inflammatory damage to muscle tissue, the effect improvement is statistically significant.

This invention belongs to the field of pharmaceutical technology, specifically relating to a FRET-based cell sensor for detecting botulinum toxin types A and E and its applications. The invention unexpectedly discovered the use of EGFP / tDimer2 as the donor and acceptor fluorophores, and SNAP-25 as the linker, to fuse and express EGFP-SANP-25-tDimer2. A cell sensor was constructed by transiently transfecting Neuro-2a cells with a plasmid expressing EGFP-SANP-25-tDimer2. This cell sensor can be applied to assess the bioactivity of materials containing BoNT / A and BoNT / E in the environment and during pharmaceutical manufacturing. It can also meet the in vitro screening requirements for botulinum toxin inhibitors and can be used for the diagnosis of BoNT / A and BoNT / E botulinum toxin poisoning.

The present application relates to botulinum toxin preparation technical field, disclose a kind of botulinum toxin type A precursor polypeptide and its preparation method and application.The botulinum toxin type A precursor polypeptide includes the light chain structure region, first connecting region and heavy chain structure region sequentially connected from N-terminal to C-terminal;The amino acid sequence of the heavy chain structure region is as shown in SEQ ID NO:3;The amino acid sequence of the first connecting region is as follows: in the connecting region amino acid sequence between natural botulinum toxin type A precursor polypeptide light chain and heavy chain, the first protease recognition sequence is inserted in the 440th position.The botulinum toxin type A precursor polypeptide of the application can improve the yield of botulinum toxin type A, and the botulinum toxin type A prepared after enzymatic activation has higher biological activity.

Provided herein are methods for improving the appearance of platysmal prominence associated with platysmal activity in a human patient by administering a composition comprising botulinum toxin type A to the platysma muscle of the human patient. Preferably, the method involves injecting the composition into the upper section of the platysma muscle below the mandibular line of the human patient and injecting the composition along one or more continuous neck bands on the neck of the human patient.

The invention provides a botulinum toxin B1 type recombinant protein and a preparation method thereof, and belongs to the technical field of biology. The amino acid sequence of the botulinum toxin B1 type recombinant protein is as shown in SEQ ID NO. 1. The invention designs a novel botulinum toxin B1 type recombinant protein with 10 His tags and Strep tags, the botulinum toxin B1 type recombinant protein can realize soluble expression in escherichia coli, the invention also provides a preparation method and a purification method of the botulinum toxin B1 type recombinant protein, and large-scale production of the botulinum toxin B1 type recombinant protein can be realized. The invention provides technical support for the application of the botulinum toxin B1.

The invention relates to optimization and transformation and application of a C1-type creotoxin light chain (BoNT / C1LC) mutant. According to the invention, a BoNT / C1LC mutant BoNT / C1alpha-51 for selectively cutting Syntaxin-1 is taken as a template, the 53th-51th amino acid of the BoNT / C1LC mutant is replaced and the enzymatic activity of the cellular level is evaluated by using a PCR site-specific mutagenesis method, and the result shows that the mutants 53W, 52I53Y and 51F52H53Y have no cutting activity on substrates SNAP-25 and Syntaxin-1, and can be applied to preparation of nervous system drug delivery targeted delivery drugs by taking full-length botulinum toxin as a carrier.

The purpose of the present invention is to provide a recombinant botulinum toxin type A light chain in which efficacy and half-life are increased to increase patient convenience and allow for customized treatment for indications, a recombinant botulinum toxin, and a composition, use, and method related thereto.

The present invention relates to a botulinum toxin type A complex, and a formulation thereof and a usage method therefor. The present invention provides the botulinum toxin type A complex, comprising an HA70 component, and an HA17 component, an HA33 component, an NINH component, and a BoNT / A1 component, wherein the botulinum toxin complex has a molecular weight of 740-790 kDa. Compared with the existing botulinum toxin complexes, the botulinum toxin complex of the present invention is smaller in molecular weight and higher in safety, and has a comparable treatment effect.

The present disclosure is to provide a recombinant botulinum toxin type A light chain, a recombinant botulinum toxin, and a composition, use, and method related thereto, which exhibit improved efficacy and half-life to be easier to use for patients and enable treatment that is customized to indications.

This invention relates to the optimization and application of C1 type botulinum toxin light chain (BoNT / C1LC) mutants. Using the selectively Syntaxin-1 cleaving mutant BoNT / C1α-51 as a template, this invention employs PCR site-directed mutagenesis to replace amino acids at positions 53-51 and evaluates enzyme activity at the cellular level. The results show that mutants 53Y, 52H53Y, 51D52H53Y, 51S52H53Y, 51G52H53Y, 51A52H53Y, and 51Y52H53Y exhibit no cleavage activity against the substrate SNAP-25, but retain cleavage activity against Syntaxin-1. Among these, 52H53Y, 51S52H53Y, and 51G52H53Y show superior Syntaxin-1 cleavage activity compared to BoNT / C1α-51. The mutants provided by this invention, which replace full-length BoNT / C1LC or are fused with a membrane-penetrating peptide and are active only for Syntaxin-1, can be used in the preparation of medical aesthetic products and related drugs that supplement clinical responses to or non-responses to botulinum toxin type A, and whether or not resistance has been developed. Mutants with Syntaxin-1 cleavage activity superior to BoNT / C1α-51 can be used in the preparation of drugs for Syntaxin-1-mediated hypersecretion diseases. Mutants such as 51F52H53Y have no cleavage activity against both SNAP-25 and Syntaxin-1 and can be used in the preparation of targeted delivery drugs for the nervous system using full-length botulinum toxin as a carrier.

This invention discloses a detection product and method for detecting botulinum toxin type A component proteins. The detection product comprises an antibody capable of specifically recognizing botulinum toxin type A component proteins. The antibody exhibits good affinity and stability, enabling its wide application in the detection of botulinum toxin type A component proteins. It can be combined with capillary immunoelectrophoresis to establish standard spectra for the detection of botulinum toxin component antibodies, providing quality control for both commercial and self-developed antibodies. Therefore, this invention offers more possibilities for establishing botulinum toxin-related detection methods and can meet the detection needs of more scenarios.

The invention provides a preparation method and application of a recombinant A-type botulinum toxin. The preparation method provided by the invention comprises the following steps: simultaneously expressing a light chain and a heavy chain of BoNTA in two expression cassettes, and autonomously assembling the light chain and the heavy chain into a soluble and active Holotoxin protein molecule in escherichia coli. According to the present invention, the recombinant BoNTA protein having the completely consistent amino acid sequence with the natural BoNTA protein can be expressed and purified, and the recombinant BoNTA can be used for expressing the truncated type or the modified type recombinant BoNTA containing one or more amino acid mutations;

The invention belongs to the technical field of biology, and particularly relates to a recombinant botulinum toxin A chimeric protein and application thereof. The recombinant A-type botulinum toxin chimeric protein is constructed by replacing only a C-terminal receptor binding domain or a whole receptor binding domain in a receptor structural domain on the basis of wild-type BoNT / A. The recombinant botulinum toxin A chimeric protein has substrate cleavage activity equivalent to that of wild toxin in vitro, can be effectively taken in Neuro-2a nerve cells and realize substrate cleavage, shows a significantly reduced toxicity level in animal bodies, and has a good application prospect. Wherein the AAAF chimeric protein has the characteristics of rapid effect taking and short action time, and is suitable for situations requiring short-term intervention of neural activity, such as short-term muscular tension regulation or local injection treatment; the AAFF chimeric protein has relatively long neuromuscular blocking duration and relatively high safety, and is suitable for long-term treatment of chronic nervous system diseases.

The invention relates to the technical field of botulinum toxin preparation, and discloses a botulinum toxin type A precursor polypeptide as well as a preparation method and application thereof. The botulinum toxin type A precursor polypeptide comprises a light chain structural region, a first connecting region and a heavy chain structural region which are sequentially connected from an N end to a C end, the amino acid sequence of the heavy chain structural region is as shown in SEQ ID NO: 3; the amino acid sequence of the first connecting region is as follows: in the amino acid sequence of the connecting region between the light chain and the heavy chain of the natural A-type botulinum toxin precursor polypeptide, a first protease recognition sequence is inserted at the 440th site. By adopting the A-type botulinum toxin precursor polypeptide disclosed by the invention, the yield of the A-type botulinum toxin can be improved, and the A-type botulinum toxin prepared after enzyme digestion and activation can have higher biological activity.

A recombinant polypeptide of the present invention comprises at least one of a recombinant receptor-binding domain of botulinum toxin type A1, a recombinant light chain domain of botulinum toxin type A, and a recombinant translocation domain of botulinum toxin type A. Also, the present invention provides a recombinant botulinum toxin type A comprising at least one of the above-described recombinant polypeptides and having increased potency, safety, or half-life compared to wild-type botulinum toxin.