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Peptide vaccine for animal and preparation method thereof

A technology for synthesizing peptide vaccines and vaccines is applied in the field of peptides of synthetic peptide vaccines for foot-and-mouth disease. synthetic effect

Inactive Publication Date: 2014-01-08
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of research on new vaccines for foot-and-mouth disease, there have been reports on genetically engineered subunit vaccines, foot-and-mouth disease virus vector vaccines, and genetically engineered vaccines for foot-and-mouth disease. use of vaccines
In addition, these vaccines are often less effective against currently circulating strains that have mutated and cannot effectively protect animals

Method used

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  • Peptide vaccine for animal and preparation method thereof
  • Peptide vaccine for animal and preparation method thereof
  • Peptide vaccine for animal and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Solid-phase synthesis of foot-and-mouth disease synthetic peptide antigen

[0034] The polypeptide antigen of the present invention can be prepared by using a fully automatic polypeptide synthesizer and Merrifield solid-phase synthesis method, in which 9-fluorenylmethyloxycarbonyl (Fmoc) modified amino acids are used, and the solid-phase carrier is Rink Amide MBHA resin. The production process usually includes solid-phase synthesis of peptide antigens, peptide lysis, antigen purification and sterilization preservation.

[0035] 1.1 Solid-phase synthesis of peptide antigens

[0036] 1.1.1 Preparation of synthetic raw materials

[0037] The sequences of the synthetic polypeptide antigens are shown in SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5 and SEQ ID NO. 6, respectively.

[0038] According to the sequence of the antigen and the synthesis scale, a suitable Fmoc modified amino acid was prepared for 1 mmol, and added to the corresponding amino a...

Embodiment 2

[0064] Example 2. Preparation of synthetic peptide vaccine

[0065] 2.1 Preparation of antigen aqueous phase

[0066] First, weigh the three polypeptide antigens synthesized according to the above-mentioned Example 1; then, dilute the synthetic peptide antigen concentration to 50μg / ml with sterile water for injection; filter the antigen solution through a filter with a pore size of 0.2μm to sterilize .

[0067] 2.2 Preparation of oil phase adjuvant

[0068] Sterilize the oil phase adjuvant at 121°C for 30 minutes, and set aside.

[0069] 2.3 Emulsification of synthetic peptide vaccine

[0070] Clean the IKA emulsification equipment 3 times with 2000ml of sterilized distilled water, and then add the oil phase to the emulsification tank at a volume ratio of 1:1 between the oil phase adjuvant and the antigen aqueous phase at 20-28°C, and start the motor After stirring at 90~150r / m for 2 minutes, slowly add the water phase antigen at the same time. After adding, stir for 30 minutes, then s...

Embodiment 3

[0071] Example 3 Efficacy test of synthetic peptide vaccine

[0072] 1. Materials and methods

[0073] 1.1 Synthetic peptide vaccine

[0074] According to the above examples, the polypeptide antigens with SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5 and SEQ ID NO. 6 were synthesized, and then prepared separately The corresponding batch numbers are: ZM62601, ZM626A02, ZM626A03, ZM626A04, ZM626A05, ZM626A06 foot-and-mouth disease synthetic peptide vaccine.

[0075] 1.2 Experimental animals

[0076] Select 32 healthy pigs of the same breed, 4 months old, about 40Kg weight, and negative for foot-and-mouth disease neutralizing antibodies.

[0077] 1.3 Virus species OR / 80MF 8

[0078] OR / 80MF 6 After passing through suckling mice for 2 generations, the virus was collected. After the suckling mice pass the poison test, they should be frozen and stored at -20°C for later use.

[0079] 1.4 Test method

[0080] For each group of vaccines, 5 healthy and susceptible shelf pigs...

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Abstract

The invention provides an aftosa synthetic peptide vaccine, more specifically to polypeptide for aftosa synthetic peptide vaccine, vaccine containing the polypeptide and a preparation method thereof. The polypeptide has amino acid sequence represented by SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 and SEQ ID NO.6. By sequencing of recent aftosa strain in China and combination of aftosa vaccine strain sequence, the invention researches variation case of main antigen site, carries out antigen site analysis and forecast assisted by computer, and carries out chemical synthesis to possible antigen site peptide segment. The invention screens polypeptide antigen through a large amount of animal tests, optimizes aftosa virus antigen site according to screening result, effectively combines T cell epitope and B cell epitope, and enhances immunization effect of the polypeptide antigen. The aftosa synthetic peptide can effectively reply antigenic variation of the aftosa virus, has good biological safety, is easy for large scale production, and has better application prospect.

Description

Technical field [0001] The present invention relates to a set of polypeptides used in foot-and-mouth disease synthetic peptide vaccines, vaccines containing these polypeptides and their preparation methods, and in particular to a polypeptide used in foot-and-mouth disease synthetic peptide vaccines, and vaccines containing the polypeptides and their preparation methods . Background technique [0002] Foot and mouth disease (FMD) is an acute, highly contact, and febrile infectious disease that occurs in cloven-hoofed animals, and is widely distributed worldwide. Because of its high infectivity and rapid spread, infection of pigs, cattle, sheep and other livestock causes the death of young animals, and the production capacity of adult animals declines sharply, which seriously endangers the development of animal husbandry and the production and supply of meat and animal products. At the same time, the market circulation and international trade of animals and animal products will be...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/09A61K39/135A61P31/14C07K1/06C07K1/04
CPCY02P20/55
Inventor 肖进齐鹏栗利芳李丙首郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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