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Canine parvovirus synthetic peptide vaccine as well as preparation method and application thereof

A technology for canine parvovirus and synthetic peptide vaccines, which is applied in the field of vaccines and their preparation, can solve problems such as purification and removal of unfavorable diseases, biological safety issues, etc., achieve good immune efficacy, easy large-scale synthesis, and no biological safety risks Effect

Active Publication Date: 2018-03-23
CHINA ANIMAL HUSBANDRY IND +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, attenuated live vaccines are commonly used in China, most of which are imported strains, which have certain biosafety problems, which are obviously not conducive to the purification and elimination of diseases

Method used

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  • Canine parvovirus synthetic peptide vaccine as well as preparation method and application thereof
  • Canine parvovirus synthetic peptide vaccine as well as preparation method and application thereof
  • Canine parvovirus synthetic peptide vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1, solid-phase synthesis of canine parvovirus synthetic peptide vaccine polypeptide

[0063] This example is the solid-phase synthesis of the polypeptide antigen of the synthetic peptide vaccine provided by the present invention. The synthetic peptide antigen of the present invention can be prepared by the Merrifield solid-phase synthesis method using a fully automatic polypeptide synthesizer, wherein amino acids modified by 9-fluorenylmethoxycarbonyl (Fmoc) are used, and the solid phase carrier is Rink Amide MBHA resin from Tianjin Nankai . The production process usually includes solid-phase synthesis of polypeptide antigens, cleavage of polypeptides, purification of antigens and sterilized storage.

[0064] 1.1 Solid-phase synthesis of synthetic peptide antigens

[0065] 1.1.1 Preparation of synthetic raw materials

[0066] The sequence of the synthetic polypeptide antigen is as follows:

[0067] Polypeptide 1 (polypeptide shown in Sequence 1 in the Seq...

Embodiment 2

[0096] Example 2, Screening of Canine Parvovirus Synthetic Peptide Vaccine Antigen Polypeptides

[0097] 1.1 Synthesis of candidate antigen polypeptides

[0098] Method is with embodiment 1.

[0099] 1.2 Screening of polypeptide antigens

[0100] Coat the 32 candidate polypeptides on the ELISA reaction plate at a concentration of 1 μg / ml, add 100 μl to each well, and place it 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 for blocking treatment at 37°C for 3 hours. Wash with wash buffer and spin dry. Add 50-fold diluted canine parvovirus antibody-positive serum (provided by Jiangxi Biopharmaceutical Factory of China Animal Husbandry Co., Ltd.) at 100 μl / well, and react in a 37°C incubator for 45 minutes. After washing, 100 μl / well of 25,000-fold diluted horseradish peroxidase-labeled rabbit anti-dog IgG antibody was added and reacted in a 37°C incubator for 45 minutes. Af...

Embodiment 3

[0104] Embodiment 3, the preparation of canine parvovirus synthetic peptide vaccine

[0105] 1.1 Preparation of antigen aqueous phase

[0106] 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.

[0107] 1.2 Preparation of oil phase adjuvant

[0108] The oil-phase adjuvant was sterilized at 121°C for 30 minutes, and set aside.

[0109] 1.3 Emulsion of synthetic peptide vaccine

[0110] 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 canine parvovirus synthetic peptide vaccine, a polypeptide composition and a vaccine. The antigen polypeptide is a polypeptide shown as asequence 1 or a sequence 2. The vaccine comprises the polypeptide or the polypeptide composition. The vaccine prepared from the polypeptide is capable of effectively coping with antigen variation ofthe canine parvovirus, is free of the problem of safety and easy to synthesize in large scale, and has good application prospects.

Description

technical field [0001] The invention relates to a polypeptide or its polypeptide polymer used for canine parvovirus synthetic peptide vaccine, vaccines containing the polypeptide or its polypeptide polymer and their preparation methods. Background technique [0002] Canine parvovirus disease is a highly contagious canine infectious disease caused by canine parvovirus (CPV) infection. The clinical manifestations are hemorrhagic enteritis and acute myocarditis, which poses a great threat to the health of dogs. Under the protection of maternal antibodies, newborn puppies are generally less likely to be infected, but puppies born 2 months old are more likely to be infected with pathogens. With the increase of immunization coverage of female dogs, the symptoms of myocarditis in newborn puppies are relatively rare in recent years, and the clinical symptoms of the current popular canine parvovirus infection are mostly hemorrhagic enteritis. Canine parvovirus was first isolated fro...

Claims

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Application Information

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IPC IPC(8): C07K14/015A61K39/23A61P31/20
CPCA61K39/12A61K2039/552C07K14/005C12N2750/14322C12N2750/14334
Inventor 向王震肖进印春生王飞李玲张晓战张国栋王楠齐鹏郑应华
Owner CHINA ANIMAL HUSBANDRY IND
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