A-type kreotoxin B cell antigen epitope peptide and use thereof

A type A botulinum toxin and epitope technology, applied in the direction of receptors/cell surface antigens/cell surface determinants, specific peptides, material inspection products, etc., can solve the problem of restricting the development of botulinum toxin and unsuitable complex epitope research , labor cycle and other issues

Inactive Publication Date: 2007-08-15
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods are time-consuming, laborious and require a long cycle, and are not suitable for

Method used

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  • A-type kreotoxin B cell antigen epitope peptide and use thereof
  • A-type kreotoxin B cell antigen epitope peptide and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Determination of Type A Botulinum Toxin Epitope Peptide

[0021] 1. Materials:

[0022] SEPHADEX G-15; SEPHAROSE affinity column; HPLC was purchased from Pharmacia;

[0023] Insight II software, SGI graphics workstation is the product of Neotrident company.

[0024] 2. Methods and results:

[0025] 1. Screening of botulinum toxin type A Hc fragment B cell epitope

[0026] In 1998, RC Stevens&DB Lacy obtained the crystal structure (PDBstructure: 3BTA, 2.3 Ȧ) of the double-chain type A botulinum toxin made up of 1285 amino acid residues through high-resolution multidimensional crystal X-ray diffraction spectrum analysis; the present invention utilizes the protein database PDB ( http: / / www.rcsb.org / pdb / home / home.do) in the three-dimensional crystal structure of botulinum toxin type A, using Insight II software and SGI graphics workstation to analyze the epitope of botulinum toxin type A B cell For analysis, select at least three consecutive amino acid residue...

Embodiment 2

[0032] Example 2 Epitope peptide immunological analysis and its application in identification of botulinum toxin serotype

[0033] 1. Materials:

[0034] Anti-botulinum toxin type A, B, E horse serum, enzyme-labeled rabbit anti-horse IgG (China Institute for the Control of Biological Products)

[0035] 2. Methods and results:

[0036] 1. ELISA analysis of specific binding of short epitope peptides to botulinum antitoxin type A horse serum

[0037] Epitope peptides 1#, 2# and 3# (10 μg / well) were coated on an enzyme-linked plate, the coating solution was NaHCO3 pH8.6, and BSA was used as a negative control, and coated overnight at 4°C. Remove the coating solution, add 100 μl 3% BSA (0.02mol / L PBS, prepared at pH 7.2) to block at 37°C for 1 hour, add anti-botulinum toxin type A horse serum (1:3000 dilution), 100 μl / well, 37°C Combine for 1 hour; wash with PBST (0.05% Tween 20); add enzyme-labeled rabbit anti-horse IgG (1:1000 dilution) 100 μl / well, incubate at room temperatur...

Embodiment 3

[0042] Example 3 Preparation of Epitope Peptide Vaccine and Its Immunoprotective Effect

[0043] 1. Materials:

[0044] Epitope peptide, prepared according to the above-mentioned embodiment;

[0045] BALB / C mice were purchased from the Experimental Animal Center of the Academy of Military Medical Sciences.

[0046] 2. Methods and results

[0047] 1. Preparation of single or multi-component epitope peptide vaccines

[0048] Single epitope peptide antigen: Epitope peptides 1#, 2# and 3# with a purity > 85% were dissolved in 0.02mol / LPBS pH7.2 respectively to prepare epitope peptide antigens with a concentration of 0.8mg / ml.

[0049] Multi-component epitope peptide antigen: epitope peptides 1#, 2# and 3# with a purity > 85%, mixed uniformly at a ratio of 1:1:1, dissolved in 0.02mol / L PBS pH7.2, and prepared as per The concentration of the epitope peptide is still the epitope peptide antigen of 0.8mg / ml.

[0050] Epitope peptide vaccines: single or multi-component epitope pep...

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Abstract

The invention discloses a cell antigen epitope peptide of A-typed botulism toxin B as core immune antigen, which is characterized by the following: protecting botulism toxin from infecting; identifying A-typed specific antibody of botulism toxin due to characteristic antigen mark; distinguishing A-type from other patterns in the serum; making the short peptide possess the same or similar function with full toxin antigen and recombinant toxin antigen without allergic reaction; overcoming the serum crossing problem among full toxin antigen or recombinant antigen; diagnosing the infection of botulism toxin to prepare new peptide vaccine; fitting for targeting and sieving botulism toxin resistant drug.

Description

technical field [0001] The present invention relates to a biotoxin B cell antigen epitope peptide, specifically relates to a botulinum toxin B cell antigen epitope peptide, and also relates to the application of the above antigen epitope peptide. Background technique [0002] Botulinum neurotoxin (Clostridium Botulinum Neurotoxins, BoNT, referred to as botulinum toxin) is an exotoxin produced by Clostridium botulinum under anaerobic conditions. Strong protein, wherein A, B, and E-type botulinum toxins are common types that cause human poisoning. Clinical observations show that A-type botulinum toxins can cause more serious symptoms and lead to higher mortality than other serotypes. [0003] The precursor of botulinum toxin type A is a non-toxic single-chain protein with a molecular weight of 150kD. It is toxic only after being cut by bacterial protease or in vitro protease into a double-chain form, that is, the light chain (L) and the heavy chain (H) pass through a Disulfid...

Claims

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Application Information

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IPC IPC(8): C07K14/705G01N33/53
Inventor 王慧史晶荫俊包士中侯晓军蔡昆
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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