Synthetic peptide vaccine for treating porcine reproductive and respiratory syndrome and preparation method thereof

A technology for respiratory syndrome and synthetic peptide vaccines, which is applied in the field of peptides for synthetic peptide vaccines, can solve the problems of short immune protection period, multiple immunizations, and strong virulence, and achieve high levels of immune response, high safety, and efficacy Good results

Inactive Publication Date: 2010-09-29
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the long-term control and eradication of diseases, both have certain shortcomings and deficiencies, such as strong virulence, loose poison, stress response of animal immunity, multiple immunizations, short immune protection period, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 : Solid Phase Synthesis of Polypeptide Antigens for Synthetic Peptide Vaccines

[0036] 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.

[0037] (1) Preparation of synthetic raw materials

[0038] Polypeptide antigens for polypeptide vaccines are synthesized, and their amino acid sequences are shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3 or SEQ ID NO.4.

[0039] Prepare appropriate Fmoc-modified amino acids according to the above peptide antigen sequence and 1 mmol synthesis scale, and add them to the corresponding Cartridge (vial containing amino acids). Also weigh th...

Embodiment 2

[0050] Example 2 : Preparation of Synthetic Peptide Vaccines

[0051] The polypeptide solution prepared in Example 1 was diluted to 200 μg / ml with water for injection, and used as the water phase; the SEPPIC MONTANIDE ISA 50V oil adjuvant was sterilized at 121°C for 30 minutes, and used as the oil phase for later use. At 22°C, according to the volume ratio of antigen water phase and sterilized 50V oil adjuvant of 1:1, first add the oil phase into the emulsification tank, stir at a slow speed of 100 rpm for 2 minutes, then slowly add the water phase, Stir for 30 minutes after the addition, then stir at a high speed of 9000 rpm for 20 minutes, then let stand for 5 minutes, and then obtain the synthetic peptide vaccine against porcine reproductive and respiratory syndrome virus after subpackaging, namely SEQID NO.1 polypeptide vaccine, SEQ ID NO. ID NO.2 polypeptide vaccine, SEQ ID NO.3 polypeptide vaccine, SEQID 4 polypeptide vaccine.

Embodiment 3

[0052] Example 3 : Efficacy Test of Synthetic Peptide Vaccines

[0053] (1) Vaccines and experimental animals

[0054] Four batches of PRRS synthetic peptide vaccine plus inactivated vaccine (purchased from China Animal Husbandry Co., Ltd. Chengdu Biopharmaceutical Factory) and adjuvant (50V oil) were set up as six groups in total, in which the inactivated vaccine was used as a positive control, the adjuvant was used as a negative control, and the experimental animals were 60 healthy pigs at the age of 40 days were tested negative for PRRSV antigen and antibody. See Table 1 for specific grouping.

[0055] Table 1 Vaccine grouping situation

[0056] group

Vaccination

Number of test animals (head)

Immunization dose (ml / head)

Group A

inactivated vaccine

10

2

Group B

SEQ ID NO.1 polypeptide vaccine

10

2

Group C

SEQ ID NO.2 polypeptide vaccine

10

2

Group D

SEQ ID NO.3 polypeptide vacci...

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Abstract

The invention provides a synthetic peptide vaccine for treating porcine reproductive and respiratory syndrome and a preparation method thereof, in particular relates to polypeptide of the synthetic peptide vaccine for treating the porcine reproductive and respiratory syndrome and a vaccine which contains the polypeptide and a method for preparing the polypeptide and the vaccine. The amino acid sequence of the polypeptide is an amino acid sequence shown in SEQ ID No.1, SEQ ID No.2, SEQ ID No.3 or SEQ ID No.4. The vaccine prepared from the polypeptide can effectively cope with the antigenic variation of a porcine reproductive and respiratory syndrome virus, ensures biological safety, is easy for large-scale synthesis and has good application prospect.

Description

technical field [0001] The present invention relates to a polypeptide used for synthesizing peptide vaccines, vaccines containing the polypeptide and preparation methods thereof, in particular to a polypeptide used for synthesizing peptide vaccines of porcine reproductive and respiratory syndrome, vaccines containing the polypeptide and its preparation method. Preparation. Background technique [0002] Porcine Reproductive and Respiratory Syndrome (PRRS), also known as PRRS, is an acute highly contagious disease caused by Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). [0003] PRRSV is a member of the genus Arterivirus in the family Arteriviridae of the order Nidoviridae. According to the differences in antigenicity, genome and pathogenicity of the virus, PRRSV can be divided into two types, namely European type (LV strain is the representative strain) and For the American type (ATCC2VR2332 strain is the representative strain), the amino acid homology between ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/08C07K1/04C07K1/06A61K39/12A61P31/14
Inventor 齐鹏肖进郭丽清巴利民栗利芳李丙首宋芳
Owner CHINA ANIMAL HUSBANDRY IND
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