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Fusion protein and application thereof to treating clostridium difficile related diseases

A technology of Clostridium difficile and fusion protein, which is applied in the field of fusion protein and its application in the treatment of Clostridium difficile-related diseases, and can solve the problems of repeated illness of patients

Active Publication Date: 2016-12-14
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

At the same time, due to the existence of spores, the spores in the recovered patients may germinate again, making the patient's condition repeated.

Method used

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  • Fusion protein and application thereof to treating clostridium difficile related diseases
  • Fusion protein and application thereof to treating clostridium difficile related diseases
  • Fusion protein and application thereof to treating clostridium difficile related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1. Different protein compositions and functional verification thereof

[0056] 1. Preparation of different protein compositions

[0057] 1. Genes encoding proteins

[0058] The amino acid sequence of TcdA is sequence 2 in the sequence listing, and the nucleotide sequence of its coding gene is sequence 1;

[0059] The amino acid sequence of TcdB is sequence 4 in the sequence listing, and the nucleotide sequence of its coding gene is sequence 3;

[0060] The amino acid sequence of BclA3 is sequence 6 in the sequence listing, and the nucleotide sequence of its coding gene is sequence 5;

[0061] The amino acid sequence of CD0873 is sequence 8 in the sequence listing, and the nucleotide sequence of its coding gene is sequence 7;

[0062] The amino acid sequence of FliD is sequence 10 in the sequence listing, and the nucleotide sequence of its coding gene is sequence 9;

[0063] The nucleotide sequences of the coding genes of TcdA, TcdB, BclA3, CD0873 and FliD we...

Embodiment 2

[0107] Embodiment 2, different fusion protein composition and application thereof

[0108] 1. Preparation of fusion protein composition

[0109] 1. Preparation of fusion protein

[0110] 1) The fusion protein TcdA-TcdB is sequentially composed of TcdA, connecting peptide (G 3 S 1 ) 3 , TcdB composition, its amino acid sequence is followed by TcdA amino acid sequence from N, (G 3 S 1 ) 3 Amino acid sequence, TcdB amino acid sequence composition; The nucleotide sequence of the coding gene of fusion protein TcdA-TcdB is followed by the nucleotide sequence of TcdA coding gene, (G 3 S 1 ) 3 Nucleotide sequence of coding gene, composition of nucleotide sequence of TcdB coding gene;

[0111] 2) The fusion protein TcdB-TcdA is sequentially composed of TcdB, connecting peptide (G 3 S 1 ) 3 , TcdA, its amino acid sequence from N is followed by TcdB amino acid sequence, (G 3 S 1 ) 3 Amino acid sequence, TcdA amino acid sequence composition; The nucleotide sequence of the c...

Embodiment 3

[0147] Embodiment 3, the immune protection of protein composition and fusion protein composition

[0148] 1. Immunoprotective properties of protein composition and fusion protein composition

[0149] 6-week-old BALB / C mice were randomly divided into 4 groups, and the following intramuscular injections were immunized respectively: (1) normal saline group; (2) the protein composition TcdA+TcdB 10 μg of embodiment 1; (3) the protein composition of embodiment 1 Protein composition BclA3+CD0873 10 μg; (4) fusion protein composition TcdB-TcdA+BclA3-CD0873 10 μg in Example 2; 100 μl each. The left leg muscle was injected on the 0th day, and the right leg muscle was injected on the 14th day; on the 28th day, 100 CFU of Clostridium difficile ATCC 43255 spores were administered orally for infection. Seven days before infection, 0.5 mg / ml cefoperazone was added to the drinking water of the mice (freshly prepared every two days, and the mice drank water freely); after 5 days of treatment...

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Abstract

The invention discloses a fusion protein and application thereof to treating clostridium difficile related diseases. The invention provides a composition composed of clostridium difficile A toxin C-terminal receptor binding domain TcdA, a clostridium difficile B toxin C-terminal receptor binding domain TcdB, a clostridium difficile spore antigen BclA3 and clostridium difficile adhesion molecules CD0873. Experiments prove that the composition immunizes mice and has good immunogenicity, and antiserum of the composition has good neutralization activity on toxin, and the composition can remarkably reduce colonization of clostridium difficile in intestinal tracts and can protect survival of the mice in clostridium difficile infection.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a fusion protein and its application in treating diseases related to Clostridium difficile. Background technique [0002] Clostridium difficile (C.difficile), Firmicutes, Clostridiales, is a Gram-positive anaerobic bacillus, widely exists in the natural environment, only infects mammals, and is transmitted through the fecal-oral route. Its spores are hardy and resistant to most disinfectants and antiseptics, allowing them to spread widely in healthcare settings. Now it has become the most important pathogenic bacteria in hospitals, causing a large amount of economic losses every year, and is the cause of more than 25% of antibiotic-associated diarrhea (Antibiotic-associated diarrhea, AAD). Mild Clostridium difficile infection can cause Clostridium difficile-associated diarrhea (CDAD), self-limited diarrhea, severe cases can develop sepsis, toxic megacolon, peritonitis and pseudomemb...

Claims

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Application Information

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IPC IPC(8): C07K19/00C07K14/33C12N15/62C12N15/31C07K16/12A61K39/08A61P31/04
CPCA61K39/08C07K14/33C07K16/1282C07K2319/00
Inventor 王慧刘坤李涛王德慧
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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