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Synthetic peptide vaccine for resisting porcine circovirus and preparation method thereof

A technology for synthesizing peptide vaccines and porcine circoviruses, applied in the field of vaccines and their preparation, can solve the problems of inability to induce cellular immune responses, weak protein immunogenicity, etc.

Active Publication Date: 2012-06-06
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Blanchard et al. (2003) compared the immune protection effects of recombinant PCV2CP and recombinant plasmids expressing CP in SPF pigs. The results showed that both recombinant protein vaccines and plasmid DNA vaccines could provide good protection, but the protective effect of recombinant CP was higher than that of recombinant Plasmids are better, and it also shows that the protein encoded by PCV2 ORF1 is weakly immunogenic and not enough to induce immune protection in the body, and it is known that recombinant protein vaccines can only stimulate the body to produce humoral immune responses, but cannot induce cellular immune responses

Method used

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  • Synthetic peptide vaccine for resisting porcine circovirus and preparation method thereof
  • Synthetic peptide vaccine for resisting porcine circovirus and preparation method thereof
  • Synthetic peptide vaccine for resisting porcine circovirus and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 : Solid Phase Synthesis of Polypeptide Antigens for Synthetic Peptide Vaccines

[0042] 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.

[0043] 1. Preparation of synthetic raw materials

[0044] The peptide antigen sequences of the synthetic peptide vaccine are respectively the amino acid sequences shown in SEQ ID NO.1, SEQ ID NO.2, SEQ INNO.3 and (SEQ ID NO.2) 2 -K-εK-CONH 2 Polymerized peptides of the indicated amino acid sequence,...

Embodiment 2

[0056] Example 2 : Formulation of Synthetic Peptide Vaccines

[0057] This example is the formulation of the synthetic peptide vaccine provided by the present invention.

[0058] Dilute the peptide solution to 50 μg / ml with water for injection to prepare the antigen aqueous phase; Seppicmontanide ISA 50V oil adjuvant was sterilized at 121°C for 30 minutes, and used 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 porcine circovirus after subpackaging, which has the amino acid sequence shown in SEQ ID NO.1. A peptide vaccine, a synthetic pe...

Embodiment 3

[0059] Example 3 : Efficacy Test of Synthetic Peptide Vaccines

[0060] In this example, the efficacy of the synthetic peptide vaccine provided by the present invention is evaluated by the following different methods.

[0061] Divide the 7-day-old healthy piglets into 5 groups, see Table 1 for the grouping information, 3 pigs in each group, inject 1ml of synthetic peptide vaccine into the muscles of the left and right sides of the neck, and inject 3 times intramuscularly, with an interval of two weeks between each time, and the third immunization Infection by oral and nasal routes after 4 weeks10 6.22 TCID 50 / ml of PCV2 tissue virus (BJ-HB strain) (gifted by China Agricultural University), each head was inoculated with 5ml.

[0062] Table 1 Grouping of experimental animals and immune challenge dose

[0063]

group

Vaccination

Number of test animals

(head)

immune dose

(ml)

Attack dose

(ml)

Group...

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Abstract

The invention provides a synthetic peptide vaccine for resisting porcine circovirus and a preparation method thereof, in particular relates to a polypeptide of the synthetic peptide vaccine for resisting porcine circovirus and a polymer thereof as well as a vaccine containing the polypeptide and a preparation method thereof. The amino acid sequence of the polypeptide is shown by SEQ ID NO.1, SEQ ID NO.2 or SEQ ID NO.3. The vaccine prepared by the polypeptide can effectively deal with antigenic variation of the porcine circovirus, has no biological safety problem, is easy to synchronize in a large scale and has favorable application prospect.

Description

technical field [0001] The invention relates to a polypeptide or its polypeptide polymer used for anti-porcine circovirus synthetic peptide vaccine, vaccines containing the polypeptide or its polypeptide polymer and their preparation methods. Background technique [0002] Porcine circovirus disease (Porcine circovirus disease, PCVD) is caused by type II circovirus. Clinically, it is an infectious disease characterized by congenital tremor of piglets, weaned piglet failure and dermatitis. Pigs can be infected and diseased, which has caused huge economic losses to the world's breeding industry. [0003] Porcine circovirus (PCV) belongs to the family Circoviridae and belongs to the genus Circovirus. Non-enveloped, single-stranded circular DNA virus. [0004] PCV is divided into two genotypes (or serotypes) according to its antigenicity and genetic composition, namely PCV1 and PCV2. PCV1 is non-pathogenic and widely exists in pigs and pig-derived cell lines; PCV2 is pathogeni...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/01C07K1/34C07K1/06C07K1/04A61K39/12A61P31/20
CPCY02P20/55
Inventor 巴利民齐鹏肖进郭丽清宋芳郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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