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Hemorrhagic bacillus coli of intestine 0157:H7 shiga toxin IIB epitope peptide and uses thereof

A kind of Escherichia coli, enterohemorrhagic technology, applied in the field of medicine and biology

Inactive Publication Date: 2008-12-03
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the structure of Stx2 and its pathogenic mechanism, drug research on Stx2 mainly focuses on Stx2 vaccines and neutralizing antibodies, but there are no Stx2 vaccines and neutralizing antibodies on the market at home and abroad.

Method used

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  • Hemorrhagic bacillus coli of intestine 0157:H7 shiga toxin IIB epitope peptide and uses thereof
  • Hemorrhagic bacillus coli of intestine 0157:H7 shiga toxin IIB epitope peptide and uses thereof
  • Hemorrhagic bacillus coli of intestine 0157:H7 shiga toxin IIB epitope peptide and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Screening and identification of B epitopes:

[0036] 1. Prediction and synthesis of epitope peptides

[0037] Retrieve the Stx2 of EHEC O157:H7 in the NCBI structural database and the crystal structure analysis diagram of Stx2 and ligand adenine (No. 1R4P and 2GA4), website: http: / / www.ncbi.nlm.nih.gov / sites / entrez? db=Structure). Based on the three-dimensional structure of the interaction between Stx2 and the ligand adenine, and based on the prediction principles of B epitope peptides exposed on the surface of the protein, containing stretches and random coils or β-turns, and peptides with a length of about 15-30 amino acids, the selected Amino acid peptides on the toxic fragment of Stx2A1, the predicted amino acid peptides were analyzed using DNAStar software for hydrophilicity (Kyte-Doolittle scheme), surface accessibility (Emini scheme), plasticity (Karp lus-Schulz scheme), secondary structure ( Garnier and Chou-Fasman scheme) and antigenicity (Jame...

Embodiment 2

[0056] Example 2 Screening of Protective B Cell Epitope Peptides by Immune Mice Challenge Test

[0057] Female Balb / c mice aged 6-8 weeks were randomly divided into 5 groups, 5 mice in each group, and 4 B cell epitope peptides (P1-KLH, P2-KLH, P3-KLH, P4- KLH) subcutaneous multi-point plus intraperitoneal injection to immunize Balb / c mice, each injected with 0.51ml, containing 100μg of antigen, and the immunization time was 0, 2, 4, 6, and 8 weeks, a total of 5 times, the first time adding Freund's complete Adjuvant Add Freund's incomplete adjuvant for the 2nd to 4th times, and intraperitoneal injection of the same dose without adjuvant for the last immunization. Blood was collected 7 days after the 2nd, 3rd, 4th, and 5th immunizations, and the immune Balb was detected by indirect ELISA. / c mouse serum titer (operating steps as before); calculate antibody positivity conversion rate, mean geometric titer (GMT) and average growth multiple. Ten days after the last immunization...

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Abstract

The invention belongs to the pharmaceutical biotechnology field, and relates to enterohemorrhagic E.coli O157:H7 Shiga toxin IIB epitope peptide and the application thereof. Four preferred polypeptides are derived from B-cell epitope of the subunit (Stx2A1) of EHEC O157:H7 Shiga toxin IIB epitope peptide. The product can be applied for preparing pharmaceuticals for the diagnosis and treatment of EHEC O157 infection and complications thereof.

Description

technical field [0001] The present invention relates to the technical field of medicine and biology, in particular to enterohaemorrhagic Escherichia coli O157:H7 Shiga toxin IIB epitope peptide and application thereof. Background technique [0002] Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is an important emerging zoonotic infectious agent, and the food poisoning caused by it has large-scale outbreaks all over the world, including China. The poisonous spinach incident that broke out in the United States in 2006 and spread to 26 states was caused by the contamination of this bacteria. From the end of 1999 to the beginning of 2000, the largest outbreak of O157:H7 infection in the world occurred in some parts of eastern my country so far. Therefore, , the infection of this bacterium has become a global public health problem. Because EHEC O157:H7 bacteria are easy to culture, strong infectivity, and diverse transmission routes, O157 bacteria are very likely to be used as...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/245C12N15/31C07K16/12A61K38/16A61K39/108A61P31/04
CPCY02A50/30
Inventor 邹全明罗萍毛旭虎曾浩石云郭刚张卫军刘璐
Owner ARMY MEDICAL UNIV
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