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Multi-epitope antigen for piglet diarrhea, vaccine, preparation method of antigen and applications of antigen and vaccine

A piglet diarrhea and multi-epitope technology, applied in the field of biotechnology applications, can solve the problems of low antibiotic efficiency and lack of cross-protection, and achieve the effect of broad protection

Pending Publication Date: 2021-09-07
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the heterogeneity of pilus adhesin expressed by ETEC, there is a lack of cross-protection between pilus adhesin antigens; in addition, the long-term use of a large number of compound antibiotics has led to the continuous generation and horizontal spread of multiple and multi-drug resistant ETEC strains, resulting in Antibiotic inefficiency and high stockpiling in animals

Method used

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  • Multi-epitope antigen for piglet diarrhea, vaccine, preparation method of antigen and applications of antigen and vaccine
  • Multi-epitope antigen for piglet diarrhea, vaccine, preparation method of antigen and applications of antigen and vaccine
  • Multi-epitope antigen for piglet diarrhea, vaccine, preparation method of antigen and applications of antigen and vaccine

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Preparation of porcine ETEC 5 Targeting Species MEFA pili adhesin protein, the following steps:

[0030] Construction and structure prediction (1) FaeG-FedF-FanC-FasA-Fim41a MEFA Protein

[0031] The main structure of the main structural sub-line using the B subunit FaeG cell epitopes predicted sites of expression of porcine ETEC fimbrial adhesin advantage F4 and F18 fimbrial adhesin subunit secondary structure FedF, and F5 pili adhesin FANC unit, the main structural subunit of pili adhesin FasA F6, F41 main structural pilus subunit B cell epitope Fim41a the online prediction.

[0032] like figure 1 The present invention is shown is the embodiment of a multi-epitope vaccine diarrhea antigen fusion constructs schematic diagram, respectively FaeG (SEQ ID NO.1), FedF (SEQ ID NO.2) as a scaffold protein, and in which 3 less immunogenic epitopes, respectively FanC (SEQ ID NO.3), FasA (SEQ ID NO.4) and Fim41a (SEQ ID NO.5) the most immunogenic epitope for alternative construct It...

Embodiment 2

[0041] Assessment FaeG-FedF-FanC-FasA-Fim41a MEFA protein as a preventive diarrhea vaccine candidate proteins:

[0042] (1) Evaluation of Immunogenicity: prokaryotic expression and purification FaeG-FedF-FanC-FasA-Fim41a MEFA protein with Freund's complete adjuvant in a 1: 1 ratio emulsified, then immunized 6-8 weeks old BALB / 16 c female mice, each mouse immunized subcutaneously 100 μl (containing 50 μg antigen). After the first immunization, booster once every two weeks apart, a total of immunity three times, in which the second and third doses of the same immunogen, but the use of adjuvant incomplete Freund's adjuvant. Another 16 mice were injected with PBS 100 μl sterile as a control group. Immunized mice were tail bled before, after the third immunization on day 14 mice were sacrificed painlessly cervical eye bled, the serum at -70 ℃ put in standby. Indirect ELISA using mouse serum corresponding pilin adhesin subunit-specific IgG antibody titers.

[0043] like Figure 4 Shown...

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Abstract

The invention discloses a multi-epitope antigen for piglet diarrhea, a vaccine, a preparation method of the antigen and applications of the antigen and the vaccine. A multi-epitope antigen fusion (MEFA) technology is utilized, and adhesin subunits FaeG and FedF of dominant expression fimbriae F4 and fimbriae F18 of ETEC strains causing piglet diarrhea are used as skeletons respectively. According to the invention, FaeG-FedF-FasA-FanC-Fim41a MEFA proteins of five fimbriae adhesin expressed by the ETEC strains capable of targeting to cause diarrhea of weaned piglets are constructed, so as to be used as effective and broad-spectrum vaccines for preventing diarrhea of newborn piglets and weaned piglets caused by ETEC infection.

Description

Technical field [0001] The present invention relates to a multi-epitope diarrhea, its preparation method and application of vaccines, and more particularly to the prediction of a B cell epitope, multiple epitope fusion antigen of bacterial etiology and construction techniques adhesin its receptor another by effectively blocking the action of high titer serum antibodies to adhesion, it belongs to the application field of biotechnology. Background technique [0002] Different multiple epitope fusion antigen (Multiepitope fusion antigen, MEFA) technology is a new vaccine computer combines biological and structural biology and technology based build platform, is derived from a strain of a different species or strains of the same expression virulence factors and (blocking) epitope, biology and the use of computers integrated into a structural biology techniques antigen, and the maximum analog epitope itself immunogenic, aims to build a force having a broad protection the new multi-epi...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/70A61K39/108A61K39/116A61P31/04
CPCC07K14/245C12N15/70A61K39/0258A61P31/04C07K2319/00A61K2039/70A61K2039/552
Inventor 段强德庞胜美朱国强吴文文
Owner YANGZHOU UNIV
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