Livestock peptide vaccine and preparation method thereof

A peptide vaccine and vaccine technology, which is applied in the field of veterinary foot-and-mouth disease chemical synthesis peptide vaccine and its preparation, can solve the problems of limited application, and achieve the effects of no biological safety, good application prospects, and easy large-scale synthesis

Inactive Publication Date: 2010-02-24
陶钰 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have confirmed that there are multiple T cell epitopes on the FMDV capsid protein that can be recognized by bovine and porcine lymphocytes, but the recognition of these T cell epitopes is obviously limited by the host MHC polymorphism, which in turn limits their use in vaccines

Method used

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  • Livestock peptide vaccine and preparation method thereof
  • Livestock peptide vaccine and preparation method thereof
  • Livestock peptide vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment I

[0036] Example I. Solid Phase Chemical Synthesis of Peptide Antigens

[0037] The preparation of the peptide antigen of the present invention can use the classic solid-phase chemical synthesis method using a fully automatic polypeptide synthesizer, using FMOC to protect amino acids, and using amino resin as the solid-phase carrier. The production process is usually divided into solid-phase synthesis of peptide antigens, cleavage of peptides, purification of antigens, and sterilized storage. Specific steps are as follows:

[0038] 1.1 Solid-phase synthesis of peptide antigens

[0039] 1.1.1 Preparation of synthetic raw materials

[0040]The sequences of the synthetic peptide antigens are shown as: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4.

[0041] Prepare suitable protected amino acids according to the above antigen sequence and synthesis scale, and add them to corresponding amino acid bottles. Also weigh the amino resin as required, put it into the reaction chamb...

Embodiment II

[0069] Example II. Proliferation assay of exogenous T cell epitopes on T cells

[0070] Test purposes:

[0071] The inventor found a powerful exogenous T cell epitope (SEQID NO: 4) on Peste des petits ruminants virus that can enhance the dominant B cell epitope at positions 130-168 on FMD VP1 through a large number of experiments, and then passed a certain The linker amino acid connects the potent exogenous T cell epitope and the dominant B cell epitope, and constructs a peptide antigen containing a helper potent exogenous T cell epitope and the main epitope gene of FMDV, which is compatible with the 130th- The antigen composed of the dominant B cell epitope at position 168 was subjected to a T cell proliferation test to examine the immunostimulatory effect of the exogenous epitope SEQ ID NO: 4 on T cells.

[0072] Test preparation:

[0073] 96-well plate, incubator, Bio-Rad microplate reader, biological safety cabinet

[0074] Preparation of 3-(4,5-dimethylthiazole-2)-2,5-...

Embodiment III

[0082] The safety test of embodiment III peptide vaccine

[0083] According to above-mentioned embodiment synthesis as the polypeptide antigen of aminoacid sequence shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, then prepare the polypeptide vaccine of anti-foot-and-mouth disease virus according to above-mentioned embodiment, every The antigen content of the batch vaccine is 60ug / ml. For each batch of vaccine, 10 guinea pigs with a body weight of 350-450 g were used for subcutaneous injection of 2 ml each; 10 mice with a body weight of 18-22 g were used for each subcutaneous injection of 0.5 ml. Continuously observe for 7 days to determine whether there is death or obvious local adverse reaction or systemic reaction caused by the injection. The observation results are shown in Table 2.

[0084] Table 2: Safety test results of peptide vaccines on guinea pigs and mice

[0085]

[0086] It can be seen from the results of this test that no death or obvious l...

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PUM

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Abstract

The invention discloses a livestock peptide vaccine resisting the infection of foot and mouth disease virus and a preparation method thereof. Susceptible animals are immunized by the peptide vaccine to resist the infection of the foot and mouth disease virus. The vaccine is combined with the capsid protein VP1 advantaged B cell epitope and the strong-effect foreign T cell epitope of the foot and mouth disease virus, has amino acid sequences presented as SEQ ID No:1, SEQ ID No:2, SEQ ID No:3 and SEQ ID No:4, can be prepared by a chemical synthetic method and a genetic engineering method, has stable product quality and good immunity effect and safety and can be used for protecting the susceptible animals from being inflected by the foot and mouth disease virus.

Description

technical field [0001] The invention relates to a veterinary foot-and-mouth disease chemically synthesized peptide vaccine and a preparation method thereof. Background technique [0002] Foot-and-mouth disease (FMD) is a highly contagious, febrile, acute infectious disease of cloven-hoofed animals, which is widely distributed in countries all over the world. It spreads rapidly and infects horses, cattle, pigs, sheep and other livestock, leading to the death of young animals and a sharp decline in the production capacity of adult animals, seriously endangering the development of animal husbandry and the production and supply of meat and animal products. Industrial production caused huge economic losses. At present, foot-and-mouth disease has been eliminated or well controlled in many countries, but in recent years, it has brought huge economic losses to the breeding industry again. In 1997, Taiwan's foot-and-mouth disease broke out, affecting more than 600 pig farms. In 200...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/135A61P31/14C07K19/00C07K14/09C07K1/06C07K1/04
Inventor 陶钰邵国虎
Owner 陶钰
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