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A mouse-derived monoclonal antibody neutralizing type A botulinum toxin and its application

A type A botulinum toxin, mouse-derived technology, applied in the fields of application, antibodies, antibacterial drugs, etc., can solve the problems of poor stability, horse serum antitoxin virus contamination, allergic reaction preparation costs, etc., and achieve the effect of clear background

Active Publication Date: 2021-06-29
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Horse serum antitoxin has risks such as virus contamination and allergic reactions, as well as disadvantages such as high preparation cost, long production period, and poor stability.

Method used

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  • A mouse-derived monoclonal antibody neutralizing type A botulinum toxin and its application
  • A mouse-derived monoclonal antibody neutralizing type A botulinum toxin and its application
  • A mouse-derived monoclonal antibody neutralizing type A botulinum toxin and its application

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Experimental program
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Effect test

Embodiment Construction

[0023] material list

[0024] Experimental animals, cells, strains: female BALB / c mice (6-8w) (Victoria), male Kunming mice (6-8w) (Victoria or the Experimental Animal Center of the Academy of Military Medical Sciences), SP2 / 0 cells (preserved in the laboratory).

[0025] Experimental reagents and consumables: 96-well ELISA plate (NEST), Protein-G affinity chromatography column (GE Company), HRP-labeled goat anti-mouse IgG (Thermo), TMB (Solebux), complete Freund's adjuvant (Sigma), incomplete Freund's adjuvant (Sigma), 1640 medium (Gibico), double antibody (Gibico), serum (Gibico-148), serum (Sigma), PEG (Sigma), mouse monoclonal antibody Ig / Subclass Identification Reagent (Boaolong), DMEM Medium (Kccell), Cell / Bacterial Total RNA Extraction Kit (Tiangen), RevertAid First Strand cDNA Synthesis Kit (Thermo), Plasmid Mini-Extraction Kit (Tiangen) , PVDF membrane (Millipore), Easysee Western Blot Kit (full gold) SDS-PAGE sample buffer (Shenggong), DMSO (Inokai), cell cultur...

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Abstract

The present invention relates to the field of biotechnology, in particular to a mouse-derived monoclonal antibody that neutralizes type A botulinum toxin and its application, and the mouse-derived monoclonal antibody that neutralizes type A botulinum toxin consists of a light chain and a heavy chain , the coding genes of CDR1, CDR2 and CDR3 in the heavy chain variable region in the heavy chain are sequentially sequence 1 from the N-terminal 71-94 nucleotides, 151-171 nucleotides and Nucleotides 286-315; the coding genes of CDR1, CDR2 and CDR3 in the light chain variable region of the light chain are sequence 3 from N-terminal 78-95 nucleotides and nucleotides in the sequence listing Nucleotides 147-155 and 259-282.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a mouse monoclonal antibody neutralizing type A botulinum toxin and its application. Background technique [0002] Botulinum toxin (BoNT) is a protein neurotoxin produced by anaerobic Clostridium botulinum (Clostridium botulinum). It is the most toxic substance known so far and can be divided into seven serotypes A-G. Types A, B, E, and F can cause botulism in humans, and type A is the most toxic and common. Botulinum toxin is listed as a Class A biological warfare agent because of its strong toxicity and ease of production. At present, food and infant botulism are still often reported, and as BoNT / A is more and more widely used in medicine and beauty, due to improper use Cases of botulism are also increasing. To sum up, we are still facing the threat of BoNT / A all the time, and its prevention and treatment should attract widespread attention. Vaccination is used to prevent BoNT / A...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/12C12N15/13A61K39/40A61P31/04
CPCA61K2039/505A61P31/04C07K16/1282C07K2317/565
Inventor 熊向华张惟材汪建华吕孙慧邱语进傅楚溪李磊孙志杰
Owner ACADEMY OF MILITARY MEDICAL SCI
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