Mouse-derived monoclonal antibody for neutralizing botulinum toxin type A and application of mouse-derived monoclonal antibody

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, long production period, and preparation cost of allergic reactions, etc., and achieve the effect of clear background

Active Publication Date: 2019-10-11
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, a...

Method used

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  • Mouse-derived monoclonal antibody for neutralizing botulinum toxin type A and application of mouse-derived monoclonal antibody
  • Mouse-derived monoclonal antibody for neutralizing botulinum toxin type A and application of mouse-derived monoclonal antibody
  • Mouse-derived monoclonal antibody for neutralizing botulinum toxin type A and application of mouse-derived monoclonal antibody

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Experimental program
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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 / subtype identification reagent (Boaolong), DMEM medium (Kccell), cell / bacterial total RNA extraction kit (Tiangen), RevertAid First Strand cDNA SynthesisKit (Thermo), plasmid mini-extraction kit (Tiangen), PVDF membrane (Millipore), Easysee Western Blot Kit (full gold), SDS-PAGE sample buffer (Shenggong), DMSO (Inokai), cell culture f...

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Abstract

The invention relates to the technical field of biology, in particular to a mouse-derived monoclonal antibody for neutralizing botulinum toxin type A and application of the mouse-derived monoclonal antibody. The mouse-derived monoclonal antibody for neutralizing the botulinum toxin type A is composed of a light chain and a heavy chain; CDR1, CDR2 and CDR3 in a heavy chain variable region in the heavy chain are sequentially amino acid residues from the 71st site to the 94th site, amino acid residues from the 151st site to the 171st site and amino acid residues from the 286th site to the 315th site from the N terminal of a sequence 2 in a sequence table; CDR1, CDR2 and CDR3 in a light chain variable region in the light chain are sequentially amino acid residues from the 78th site to the 95thsite, amino acid residues from the 147th site to the 155th site and amino acid residues from the 259th site to the 282nd site from the N terminal of a sequence 4 in the sequence table.

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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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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