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CN-121499819-B - Detection product and method for detecting A-type botulinum toxin component protein

CN121499819BCN 121499819 BCN121499819 BCN 121499819BCN-121499819-B

Abstract

The invention discloses a detection product and a detection method for detecting A-type botulinum toxin component proteins. The detection products of the present invention comprise antibodies capable of specifically recognizing botulinum toxin type A component proteins. The antibody provided by the invention has good affinity and stability, can be widely applied to detection of A-type botulinum toxin component protein, can be combined with a capillary immunoelectrophoresis method to establish a standard spectrum for detecting the botulinum toxin component antibody, and provides a quality control means for commercialized and self-ground antibodies, so that the invention provides more possibilities for establishing a botulinum toxin related detection method, and can meet detection requirements of more scenes.

Inventors

  • MA YAFENG
  • LU JIAN
  • WANG YIZHI
  • ZHU LI
  • ZHANG RUOHUI

Assignees

  • 兰州生物技术开发有限公司

Dates

Publication Date
20260505
Application Date
20260109

Claims (10)

  1. 1. An assay product for detecting botulinum toxin type a component protein, wherein the assay product is for detecting hemagglutinin-70, the assay product comprising an antibody or antigen-binding fragment thereof comprising heavy chain CDRs 1-3 having the amino acid sequences shown in SEQ ID nos. 1-3 and light chain CDRs 1-3 having the amino acid sequences shown in SEQ ID nos. 4-6.
  2. 2. A method of detecting botulinum toxin type a hemagglutinin-70, wherein the method is a non-disease diagnostic method comprising the steps of: (a) Providing a sample to be tested; (b) Contacting the test sample with an antibody or antigen-binding fragment thereof comprising heavy chain CDRs 1-3 having the amino acid sequences shown in SEQ ID nos. 1-3 and light chain CDRs 1-3 having the amino acid sequences shown in SEQ ID nos. 4-6 for a time and under conditions sufficient to form an antibody or antigen-binding fragment/antigen complex thereof; (c) Detecting binding of said antibody or antigen binding fragment thereof to said sample to be tested, or detecting the presence of said antibody or antigen binding fragment/antigen complex thereof, to determine the presence of hemagglutinin-70 in said sample or to determine the amount of hemagglutinin-70 in said sample.
  3. 3. An antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof is capable of targeting botulinum toxin A hemagglutinin-70, and wherein the antibody or antigen-binding fragment thereof comprises heavy chain CDR1-3 having the amino acid sequence shown in SEQ ID NO.1-3 and light chain CDR1-3 having the amino acid sequence shown in SEQ ID NO. 4-6.
  4. 4. An antibody or antigen-binding fragment thereof according to claim 3, wherein the antibody or antigen-binding fragment thereof has a heavy chain with the amino acid sequence shown in SEQ ID No.7 and a light chain with the amino acid sequence shown in SEQ ID No. 8.
  5. 5. An antibody or antigen-binding fragment thereof according to claim 3, wherein the antibody is selected from the group consisting of monoclonal antibodies and the antigen-binding fragment is selected from the group consisting of Fab, fab ', F (ab') 2 , or scFv.
  6. 6. A nucleic acid molecule comprising a nucleotide sequence encoding an antibody or antigen-binding fragment thereof according to any one of claims 3-5.
  7. 7. A vector molecule, characterized in that it comprises a nucleic acid molecule according to claim 6.
  8. 8. A host cell comprising a nucleic acid molecule according to claim 6 or a vector molecule according to claim 7.
  9. 9. The method of producing an antibody or antigen-binding fragment thereof according to any one of claims 3 to 5, wherein the antibody or antigen-binding fragment thereof is produced synthetically or by genetic engineering.
  10. 10. Use of an agent for the preparation of a product for the detection of botulinum toxin type a hemagglutinin-70, characterized in that the agent comprises an antibody or an antigen-binding fragment thereof according to any one of claims 3 to 5.

Description

Detection product and method for detecting A-type botulinum toxin component protein Technical Field The invention belongs to the field of botulinum toxin detection, and particularly relates to a detection product and a detection method for detecting A-type botulinum toxin component proteins. Background Botulinum toxin (botulinumtoxin, boNT) is a protein neurotoxin produced by clostridium botulinum (Clostridium botulinum) and has eight serotypes a-H. The botulinum toxin type A complex protein consists of neurotoxin, non-toxic non-hemagglutinin and hemagglutinin components, and most of the toxin protein components have certain immunogenicity and can generate corresponding antibodies. The problems in the detection of botulinum toxin and corresponding antibodies are that firstly, in the development and production of botulinum toxin related products, antibodies against toxin complex proteins or neurotoxin proteins are inevitably used, whether from commercial sources or self-developed antibodies, standards are lacking, effective quality control means are lacking, the actual specificity of antibodies and binding activity against component proteins are difficult to verify, secondly, commercial antibodies against the complete set of botulinum toxin components are lacking in the market, self-developed antibodies lack commercial specific component proteins as antigens, and effective purification means against the natural toxin specific component proteins are temporarily absent, and thirdly, the deletion of antibodies and specific component proteins simultaneously hinders the establishment of some botulinum toxin related detection methods, such as monitoring the integrity and purity of toxin complex proteins by immunological methods, identification of different component botulinum toxin products, verification of botulinum toxin component expression, and the like. Thus, there is a need for detection products and methods that can detect botulinum toxin component proteins. Disclosure of Invention In order to solve at least part of the technical problems in the prior art, the invention provides a detection product and a detection method for detecting a botulinum toxin type A component protein. Specifically, the present invention includes the following. In a first aspect of the present invention there is provided an assay product for detecting botulinum toxin type A component protein, wherein the assay product is for detecting hemagglutinin-70, the assay product comprising an antibody or antigen-binding fragment thereof comprising heavy chain CDR1-3 of the amino acid sequence shown in SEQ ID NO.1-3 and/or light chain CDR1-3 of the amino acid sequence shown in SEQ ID NO. 4-6. In certain embodiments, the detection product according to the present invention, wherein the detection product comprises a kit, a test strip or a protein chip. In a second aspect of the invention, there is provided a method of detecting botulinum toxin type A hemagglutinin-70, comprising the steps of: (a) Providing a sample to be tested; (b) Contacting the test sample with an antibody or antigen-binding fragment thereof comprising heavy chain CDRs 1-3 having the amino acid sequences shown in SEQ ID nos. 1-3 and/or light chain CDRs 1-3 having the amino acid sequences shown in SEQ ID nos. 4-6 for a time and under conditions sufficient to form an antibody or antigen-binding fragment/antigen complex thereof; (c) Detecting binding of said antibody or antigen binding fragment thereof to said sample to be tested, or detecting the presence of said antibody or antigen binding fragment/antigen complex thereof, to determine the presence of hemagglutinin-70 in said sample or to determine the amount of hemagglutinin-70 in said sample. In a third aspect of the invention, there is provided an antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof is capable of targeting botulinum toxin type A hemagglutinin-70, the antibody or antigen-binding fragment thereof comprising heavy chain CDR1-3 having the amino acid sequence shown in SEQ ID NO.1-3 and/or light chain CDR1-3 having the amino acid sequence shown in SEQ ID NO. 4-6. In certain embodiments, an antibody or antigen-binding fragment thereof according to the invention, wherein the antibody or antigen-binding fragment thereof has any one of the amino acid sequences shown in (I) - (III): (I) A heavy chain amino acid sequence shown in SEQ ID No.7 and/or a light chain amino acid sequence shown in SEQ ID No. 8; (II) an amino acid sequence having at least 90% homology with the amino acid sequence shown in (I) and having the same function; (III) an amino acid sequence having the same function as the amino acid sequence shown in (I) or (II) obtained by modifying, substituting, deleting or adding one or more than one amino acid. In certain embodiments, an antibody or antigen-binding fragment thereof according to the present invention, wherein the antibody comprises a monoclonal