Acinetobacter baumannii omp22 recombinant multi-antigen epitope polypeptide and its application

A technology of Acinetobacter baumannii and epitope polypeptide, applied in A.bOmp22 recombinant multi-antigen epitope polypeptide and application field, can solve problems such as toxicity and harm, and achieve the effect of avoiding toxic effect, reducing infection and epidemic

Active Publication Date: 2022-05-17
THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Omp22 is both involved A.b The key protein in the metabolic process has a certain degree of toxicity. If the full-length protein is used alone as a vaccine antigen to immunize the host, it will be harmful to a certain extent.

Method used

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  • Acinetobacter baumannii omp22 recombinant multi-antigen epitope polypeptide and its application
  • Acinetobacter baumannii omp22 recombinant multi-antigen epitope polypeptide and its application
  • Acinetobacter baumannii omp22 recombinant multi-antigen epitope polypeptide and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] A.b Construction and screening of polypeptide sequences of B cell antigen epitope and T cell antigen epitope of Omp22 protein, and chemical synthesis of recombinant multi-antigen epitope polypeptide rOmp22. Specific steps are as follows:

[0046] The amino acid sequences of 4 B-cell epitopes and 4 T-cell epitopes of Acinetobacter baumannii Omp22 protein are:

[0047] A.b Omp22 175-182: GKGVPSSR;

[0048] A.b Omp22 158-172: NIPLSQARAQSVKNY;

[0049] A.b Omp22 125-135: YATLDKVAQTL;

[0050] A.b Omp22 102-108: SVQLIMP;

[0051] A.b Omp22 112-122: TFDTNKSNIKP;

[0052] A.b Omp22 153-164: GNDSINIPLSQ;

[0053] A.b Omp22 178-188: VPSSRIDAQGY;

[0054] A.b Omp22 204-215: EQNRRVEISIY.

[0055] Entrust Nanjing KingScript Biotechnology Co., Ltd. to chemically synthesize, with a purity of more than 85%, and couple BSA molecules to the C-terminus of each epitope short peptide;

[0056] A. Construct the recombinant plasmid pET28a-omp22, transform the recomb...

Embodiment 2

[0070] For the detection of the immune effect of the recombinant multi-antigen epitope polypeptide rOmp22, the specific implementation steps are as follows:

[0071] Female BALB / c mice at 6-8 weeks were randomly divided into 4 groups, 6 in each group, respectively high-dose rOmp22 group (40ug / mouse), low-dose rOmp22 group (10ug / mouse), and Fu Adjuvant 1:1 was mixed and fully emulsified. A normal saline + adjuvant group and a normal saline control group were set up, with a total volume of 200ul. They were injected subcutaneously for immunization. Complete Freund's adjuvant was used for the first immunization, and Freund's non-toxic adjuvant was used for the last two immunizations. Complete adjuvant, once every two weeks, a total of three immunizations. One week after the last immunization, the mouse serum was collected and the spleen was isolated.

[0072] The ELISA method is used to detect the expression level of rOmp22-specific IgG in mouse serum. The steps are as follows: 2...

Embodiment 3

[0075] Recombinant multi-antigen epitope polypeptide rOmp22 pair A.b The resulting mouse pneumonia has an immune protective effect, and the specific implementation steps are as follows:

[0076] 6-8 weeks female BALB / c mice were randomly divided into 4 groups (10 mice in each group), respectively high-dose rOmp22 group (40ug / mouse), low-dose rOmp22 group (10ug / mouse), and Freund's adjuvant was mixed and fully emulsified at 1:1, and a normal saline + adjuvant group and a normal saline control group were set up, with a total volume of 200ul, and were injected subcutaneously for immunization. Freund's complete adjuvant was used for the first immunization, and Freund's for the last two immunizations. Incomplete adjuvant, once every two weeks, a total of three immunizations.

[0077] 2 weeks after the last immunization, 50ul half-lethal dose of A.b Standard strain ATCC 19606 (1×10 8 CFU, containing 5% porcine mucin), to establish a mouse model of acute pneumonia.

[0078] 24 ...

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Abstract

The present invention belongs to the application of Acinetobacter baumannii Omp22; specifically relates to Acinetobacter baumannii Omp22 recombinant multi-antigen epitope polypeptide and its application; A.b The highly conserved and highly immunogenic Omp22 protein was used as the target antigen, and the dominant B cell epitopes and T cell epitopes of Omp22 protein were screened by bioinformatics and immunology methods, and each dominant antigenic epitope was identified by 6-aminocaproic acid. The polypeptides are connected in series, and the recombinant multi-antigen epitope polypeptide rOmp22 is chemically synthesized, which not only retains A.b Omp22 protein immunogenicity, and avoid the virulence of the original structural protein, to develop a safe and effective anti- A.b New vaccines for bacterial infections, to reduce A.b Infection and prevalence in hospitals provides a new tool. Animal experiments show that the recombinant vaccine can induce significant cellular and humoral immune responses in the body, has immune protection against Acinetobacter baumannii infection, and is highly safe and convenient to use.

Description

technical field [0001] The invention belongs to the application of the outer membrane protein Omp22 of Acinetobacter baumannii, in particular to a method for treating Acinetobacter baumannii ( Acinetobacter baumannii, A.b. ) infection with immunoprotective effect A.b Omp22 recombinant multi-antigen epitope polypeptide and its application. Background technique [0002] Acinetobacter baumannii ( Acinetobacter baumannii, A.b. ) is a non-fermenting gram-negative bacillus, which is an opportunistic pathogen. It is widely distributed in the hospital environment and can survive for a long time. It can cause infections in the respiratory system, digestive system, urinary system, and central system, and even bloodstream infections. Morbidity and mortality And the drug resistance rate is in the forefront of Gram-negative bacilli infection. A.b The mechanism of drug resistance is complex, including the generation of various inactivated enzymes, the loss of outer membrane proteins,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/21C07K16/12C12N15/70A61K39/104A61P31/04
CPCC07K14/212C07K16/1217C12N15/70A61K39/1045A61P31/04
Inventor 冯旰珠杜兴冉姜明子邓凯丽陈思遐林少清黄榆珍沈自燕
Owner THE SECOND AFFILIATED HOSPITAL OF NANJING MEDICAL UNIV
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