Porcine circovirus synthetic peptide vaccine and preparation method and application thereof

A technique for synthesizing peptide vaccines and porcine circoviruses, which is applied in the field of medicine, can solve the problems of difficulty in uniform quality between batches, high prices, and limited control effects of porcine circovirus diseases, and achieve improved safety and specificity and reduced production costs. Inexpensive, reliable vaccine quality results

Active Publication Date: 2017-02-08
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 circovirus synthetic peptide vaccine and preparation method and application thereof
  • Porcine circovirus synthetic peptide vaccine and preparation method and application thereof
  • Porcine circovirus synthetic peptide vaccine and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1, solid-phase synthesis of porcine circovirus synthetic peptide vaccine polypeptide

[0072] The present invention studies the variation of the main antigenic site of porcine circovirus type 2 by determining the sequence of the domestic porcine circovirus type 2 newly popular strain 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 high l...

Embodiment 2

[0112] Example 2, Screening of Porcine Circovirus Synthetic Peptide Vaccine Antigen Polypeptides

[0113] 1.1 Synthesis of candidate antigen polypeptides

[0114] The synthetic method is the same as in Example 1.

[0115] 1.2 Screening of antigenic polypeptides

[0116] The inventor of the present invention designed and screened the antigenic polypeptides through a large number of literature reviews, and after multiple screenings among tens of thousands of polypeptides, and repeatedly modified and optimized some sites, finally obtained 36 polypeptides as candidates for polypeptide vaccines .

[0117] 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 washing buffer and spin dry. Add 60-fold diluted por...

Embodiment 3

[0122] Embodiment 3, the preparation of porcine circovirus synthetic peptide vaccine

[0123] 1.1 Preparation of antigen aqueous phase

[0124] Weigh the synthetic peptide antigens shown in Polypeptide 1, Polypeptide 2, Polypeptide 3 or Polypeptide 4 prepared according to Example 1, mix them according to the molar ratio of the antigens at 1:1:1:1, and then use sterile water for injection to dissolve the synthetic peptide antigens The total antigen concentration was diluted to 100 μg / ml. The resulting antigen solution was filtered through a filter with a pore size of 0.2 μm and sterilized.

[0125] 1.2 Preparation of oil phase adjuvant

[0126] Sterilize the oil phase adjuvant at 50V at 121°C for 30min, and set aside.

[0127] 1.3 Emulsion of synthetic peptide vaccine

[0128] 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 ...

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Abstract

The invention provides antigen polypeptide, a polypeptide composition and a vaccine for preparing a porcine circovirus synthetic peptide vaccine. The antigen polypeptide is polypeptide shown in the sequence 1, sequence 2, sequence 3 or sequence 4. The synthetic peptide vaccine comprises polypeptide or a polypeptide composition. The provided porcine circovirus synthetic peptide vaccine is excellent in immune effect and doesn't cause problems of fever and redness existing in conventional vaccine. The vaccine can effectively overcome the problems including antigenic variation of porcine circovirus and absence of biological safety, is easy for massive synthesis, and has bright application prospects. The invention further provides a preparation method of polypeptide and a polypeptide vaccine and a pharmaceutical application thereof.

Description

technical field [0001] The invention relates to the technical field of medicine, and in particular, the invention relates to a porcine circular synthetic peptide vaccine and a preparation method thereof. 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 PCV2 infects the body, it invade...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/01A61K39/12A61P31/20
CPCA61K39/00A61K2039/552A61P31/20C07K14/005C12N2750/10022C12N2750/10034
Inventor 栗利芳贾淼王楠刘新月齐鹏肖进赵洪涛张欣张艳宾张君
Owner CHINA ANIMAL HUSBANDRY IND
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