Swine fever synthesized peptide vaccine and preparation method thereof

A technique for synthesizing peptide vaccines and vaccines, which is applied in the field of peptides of swine fever synthetic peptide vaccines, can solve the problems of loss of efficacy of biohazardous original vaccines, and achieve the effects of good application prospects, high safety, and easy large-scale synthesis

Inactive Publication Date: 2014-06-04
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Traditional vaccines are made by inactivating or attenuating pathogens, and there are problems such as biological hazards and genetic variation that cause the original vaccine to lose efficacy

Method used

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  • Swine fever synthesized peptide vaccine and preparation method thereof
  • Swine fever synthesized peptide vaccine and preparation method thereof
  • Swine fever synthesized peptide vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 : Solid Phase Synthesis of Polypeptide Antigens for Synthetic Peptide Vaccines

[0033]This example is the solid-phase synthesis of the polypeptide antigen of the synthetic peptide vaccine provided by the present invention. The polypeptide antigen of the present invention can be prepared by ABI 433A automatic polypeptide synthesizer by Merrifield solid-phase synthesis method, wherein 9-fluorenylmethoxycarbonyl (Fmoc) modified amino acid is used, and the solid phase carrier is Rink Amide MBHA resin. The production process usually includes solid-phase synthesis of polypeptide antigens, cleavage of polypeptides, purification of antigens and sterilized storage.

[0034] 1. Preparation of synthetic raw materials

[0035] The polypeptide antigen sequences of the synthetic peptide vaccine are the amino acid sequences shown in SEQ ID NO.1, SEQ ID NO.2, SEQ IN NO.3 and SEQ ID NO.4 respectively.

[0036] Prepare appropriate Fmoc-modified amino acids according to the...

Embodiment 2

[0047] Example 2 : Formulation of Synthetic Peptide Vaccines

[0048] This example is the formulation of the synthetic peptide vaccine provided by the present invention.

[0049] Dilute the peptide solution to 50 μg / ml with water for injection to prepare the antigen aqueous phase; sterilize the SEPPIC MONTANIDE ISA 50V oil adjuvant at 121°C for 30 minutes, and use it as the oil phase for later use. Under the condition of 22°C, according to the volume ratio of the antigen water phase and the sterilized oil phase 50V as 1:1, the oil phase was first added to the emulsification tank, stirred at a slow speed of 100 rpm for 2 minutes, and then the water phase was slowly added, and the Stir for 30 minutes after completion, then stir at a high speed of 9000 rpm for 20 minutes, then let stand for 5 minutes, and then obtain a synthetic peptide vaccine against avian influenza virus after subpackaging, i.e. a synthetic peptide vaccine with the amino acid sequence shown in SEQ ID NO.1 ,...

Embodiment 3

[0050] Example 3 : Efficacy Test of Synthetic Peptide Vaccines

[0051] This example provides the efficacy test of the synthetic peptide vaccine of the present invention, which is detailed as follows.

[0052] 1. Test method: The efficacy of the swine fever synthetic peptide vaccine prepared in Example 2 for immunizing pig serum was detected by blocking ELISA.

[0053] The four batches of synthetic peptide vaccines were divided into four groups in total. The test animals were 16 healthy pigs that were negative for CSF antigen and antibody at the age of 40 days. See Table 1 for the specific grouping conditions. Before immunization and 21 days after the second immunization, the blood was collected from the ear vein to separate the serum, which was used as the test serum.

[0054] Table 1 Vaccine grouping situation

[0055] group Vaccination Number of test animals (head) Immunization dose (ml / head) Group 1 SEQ ID NO.1 polypeptide vaccine 4 2 2 ...

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Abstract

The invention provides a swine fever synthesized peptide vaccine and a preparation method thereof. The invention specifically relates to polypeptide used for synthesizing the swine fever synthesized peptide vaccine, a vaccine containing the polypeptide, and a preparation method thereof. The amino acid sequence of the polypeptide is represented by SEQ ID No.1, SEQ ID No.2, SEQ ID No.3, or SEQ ID No.4. With the vaccine prepared from the polypeptide, antigen variations of swine fever viruses can be effectively coped with, and a bio-security problem is avoided. The polypeptide vaccine is suitable for large-scale synthesizing, and has a good application prospect.

Description

technical field [0001] The invention relates to a polypeptide used for a synthetic peptide vaccine of swine fever, a vaccine containing the polypeptide and a preparation method thereof. Background technique [0002] Classical Swine Fever (CSF) is an important viral infectious disease that seriously endangers the swine industry. Its main manifestations are characteristic pathological changes, visceral hemorrhage, infarction and necrosis, and the mortality rate is as high as 80% to 90%. The world (except North America and Oceania), causing significant economic losses to the pig industry. The International Organization for Animal Health has listed swine fever as a severe infectious disease of Class A. Swine fever only infects pigs under natural conditions. Pigs and wild boars of different ages, sexes, and breeds are susceptible and can occur throughout the year. In addition, sows infected with sick and attenuated strains can also vertically infect fetuses through the placenta...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/18C07K1/34C07K1/06C07K1/04A61K39/187A61P31/14
CPCA61K39/00C07K14/005C12N2770/24322C12N2770/24334
Inventor 齐鹏宋芳郭丽清肖进巴利民郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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