Porcine circular synthetic peptide vaccine and its preparation method and application

A technology for synthesizing peptide vaccines and porcine circovirus, which is applied in the field of medicine, can solve the problems of limited control effect of porcine circovirus disease, difficulty in culturing inactivated vaccine viruses, and low production yield of inactivated vaccines, and achieves good immune efficacy, The effect of reliable vaccine quality and low production cost

Active Publication Date: 2020-08-25
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These two vaccines have played an important role in the prevention and control of PCV2 infection in my country, but at the same time there are also difficulties such as the difficulty in cultivating the inactivated vaccine virus, the difficulty in unifying the quality between batches during production, and the need for further improvement in the production process, which makes the production of inactivated vaccines insufficient. High, and there is also the risk of spreading poison
Although the baculovirus vector vaccine can ensure the stability and consistency of the vaccine and easily distinguish the immunized animals from the infected animals, it is expensive, and many pig farms cannot purchase in large quantities, so the control effect on porcine circovirus disease in my country limited

Method used

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  • Porcine circular synthetic peptide vaccine and its preparation method and application
  • Porcine circular synthetic peptide vaccine and its preparation method and application
  • Porcine circular synthetic peptide vaccine and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1, solid-phase synthesis of porcine circovirus synthetic peptide vaccine polypeptide

[0063] The present invention studies the variation of the main antigenic site of porcine circovirus type 2 by determining the sequence of the newly popular strain of porcine circovirus type 2 in China and combining the sequence of the porcine circovirus type 2 vaccine strain, and aims at the main mutated amino acid site Statistical variation frequency, combined with computer-aided analysis and prediction of FMD antigenic sites, chemical synthesis of possible antigenic site peptides, that is, according to the statistical variation frequency of variable sites, use different Amino acids to obtain a variety of candidate polypeptide antigens covering all possible mutation sites. Furthermore, through a large number of animal experiments, these candidate polypeptide antigens were screened to obtain a polypeptide antigen that can cause an immune response in animals, and has a hig...

Embodiment 2

[0096] Example 2, Screening of Porcine Circovirus Synthetic Peptide Vaccine Antigen Polypeptides

[0097] 1.1 Synthesis of candidate antigen polypeptides

[0098] Synthetic method is the same as embodiment 1.

[0099] 1.2 Screening of antigenic polypeptides

[0100] The 36 candidate antigen polypeptides were respectively coated on the ELISA reaction plate at a concentration of 1 μg / ml, and placed overnight at 4-8°C to fully combine the polypeptide antigens with the enzyme-linked reaction plate. Then, 300 μl / well of blocking solution was added to block at 37°C for 2-3 hours. Wash with wash buffer and spin dry. Add 60-fold diluted porcine circovirus type 2 antibody-positive serum (provided by China Animal Husbandry Co., Ltd. Chengdu Pharmaceutical Equipment Factory) at 100 μl / well, and react in a 37°C incubator or water bath for 30 minutes. After washing, 100 μl / well of 10,000-fold diluted horseradish peroxidase-labeled rabbit anti-pig IgG antibody (Sigma, product number A...

Embodiment 3

[0105] Embodiment 3, the preparation of porcine circovirus synthetic peptide vaccine

[0106] 1.1 Preparation of antigen aqueous phase

[0107] Weigh the synthetic peptide antigens shown in polypeptide 1 and polypeptide 2 prepared according to Example 1, mix them according to the molar ratio of the antigens at 1:1, and then dilute the total concentration of the synthetic peptide antigens to 100 μg / ml with sterile water for injection. The resulting antigen solution was filtered through a filter with a pore size of 0.2 μm and sterilized.

[0108] 1.2 Preparation of oil phase adjuvant

[0109] Sterilize the oil phase adjuvant at 50V at 121°C for 30min, and set aside.

[0110] 1.3 Emulsion of synthetic peptide vaccine

[0111] Clean the IKA emulsification equipment with 2000ml of sterilized distilled water for 3 times, then put the oil phase into the emulsification tank at 20-28°C according to the volume ratio of oil phase adjuvant and antigen water phase at 1:1, and start th...

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Abstract

The invention discloses an antigen polypeptide for preparing a porcine circovirus synthetic peptide vaccine, a polypeptide composition and a vaccine. The antigen polypeptide is a polypeptide with sequence 1 or sequence 2. The vaccine comprises the polypeptide or the polypeptide composition. The porcine circovirus synthetic peptide vaccine provided by the invention has good immune potency, and does not induce fever, swelling and other problems like traditional vaccines. Therefore, the vaccine of the invention can effectively deal with the antigenic variation of porcine circovirus, has good biosafety, is easy for large-scale synthesis, and has good application prospects. The invention also provides a preparation method and pharmaceutical applications of the polypeptide and the polypeptide vaccine.

Description

technical field [0001] The present invention relates to the technical field of medicine, and in particular, the present invention relates to a porcine circular synthetic peptide vaccine as well as its preparation method and application. Background technique [0002] Porcine circovirus-associated disease (PCVAD) refers to a series of syndromes caused by PCV2 or diseases related to PCV2 infection. According to the difference in PCV antigenicity and genetic composition, it is divided into two genotypes (serotypes). The porcine circovirus associated with clinical disease is named porcine circovirus type 2 (PCV2). The circovirus is named porcine circovirus type 1 (PCV1). Porcine circovirus is the smallest animal virus discovered so far, with a diameter of 17nm, icosahedral symmetry, and no envelope. The viral genome consists of single-stranded negative-strand closed circular DNA, of which PCV1 contains 1759 nucleotides and PCV2 contains 1767 or 1768 nucleotides. [0003] After...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/01A61K39/12A61P31/20
CPCA61K39/12A61K2039/552C07K14/005C12N2750/10022C12N2750/10034
Inventor 栗利芳贾淼刘新月王楠肖进齐鹏宋芳巴利民
Owner CHINA ANIMAL HUSBANDRY IND
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