A kind of bird flu synthetic peptide vaccine and preparation method thereof

A technology for synthesizing peptide vaccines and avian influenza, applied in the field of medicine, can solve problems such as the loss of efficacy of biohazardous original vaccines, and achieve the effect of high level of immune response

Active Publication Date: 2021-02-09
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Traditional vaccines are made by inactivating or attenuating pathogens, and there are problems such as biological hazards and genetic variation that cause the original vaccine to lose efficacy

Method used

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  • A kind of bird flu synthetic peptide vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: the solid-phase synthesis of synthetic peptide vaccine polypeptide antigen

[0036] 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 a Merrifield solid-phase synthesis method using a fully automatic polypeptide synthesizer, wherein amino acids modified with 9-fluorenylmethoxycarbonyl (Fmoc) are 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] The polypeptide antigen sequence of the synthetic peptide vaccine is the amino acid sequence shown in sequence 1.

[0039] Prepare suitable Fmoc-modified amino acids according to the above polypeptide antigen sequence and the synthesi...

Embodiment 2

[0050] Embodiment 2: the preparation of synthetic peptide vaccine

[0051] This example is the formulation of the synthetic peptide vaccine provided by the present invention.

[0052] Dilute the peptide solution to 50pg / ml with water for injection to prepare the antigen aqueous phase; sterilize the SEPPIC MONTANIDE ISA61 oil adjuvant at 121°C for 30 minutes, and use it as the oil phase for later use. Under the condition of 20-25°C, according to the volume ratio of antigen water phase and sterilized 61 oil adjuvant as 1:1, first add the oil phase into the emulsification tank, and stir slowly at 80-100 rpm for 2-5 Minutes, slowly add the water phase, stir for 20-30 minutes after the addition, and then stir at a high speed of 8000-10000 rpm for 15-30 minutes, then let stand for 5 minutes, and obtain the synthetic peptide vaccine against avian influenza virus after sub-packaging.

Embodiment 3

[0053] Embodiment 3: the immune test of synthetic peptide vaccine

[0054] In this example, an immune test is carried out on the synthetic peptide vaccine provided by the present invention, and the immune effect of the synthetic peptide vaccine is evaluated using a serological test.

[0055] The 4-5 week-old SPF chickens were randomly divided into two groups, 10 SPF chickens in the test vaccine group, and 5 SPF chickens in the control group. The specific grouping conditions are shown in Table 1.

[0056] Table 1 Vaccine grouping and number of animals required

[0057] group vaccine SPF chicken (only) Group A experimental vaccine group 10 Group B saline control group 5

[0058] The neck of each SPF chicken in the test vaccine group was subcutaneously injected with 0.3 ml of the synthetic peptide vaccine against avian influenza virus prepared in Example 2, and the neck of the SPF chicken of the control group was subcutaneously injected with 0.3 ...

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Abstract

The invention provides a polypeptide antigen for preparing a bird flu synthetic peptide vaccine, a vaccine containing the polypeptide antigen and a preparation method thereof. Polypeptide antigens include the polypeptides shown in Sequence 1. The avian influenza synthetic peptide vaccine provided by the invention has good immune efficacy, and the antigen of the vaccine is a chemically synthesized polypeptide, does not contain avian influenza virus nucleic acid particles, and has high safety. Therefore, the vaccine of the invention can effectively deal with the antigenic variation of the avian influenza virus, has no biological safety, is easy to synthesize on a large scale, and has good application prospects.

Description

technical field [0001] The invention belongs to the technical field of medicine, in particular, the invention relates to a novel avian influenza synthetic peptide vaccine and a preparation method thereof. Background technique [0002] Avian influenza (Avian influenza, AI) is an infectious disease caused by avian influenza virus (AIV). animal diseases. AIV is a type A influenza virus belonging to the genus Influenzavirus of the Orthoquaviridae family. Its genome is composed of 8 segmented single-stranded negative-sense RNAs, which encode 8 proteins with different functions: polymerase proteins PB1, PB2, and PA; Hemagglutinin protein HA; Nucleocapsid protein NP; Neuraminidase protein NA; Matrix protein M1 and M2, and nonstructural protein NS1 and NS2, wherein, hemagglutinin (HA), neuraminidase (NA) and M2 proteins are three surface antigen molecules of avian influenza virus, and antibodies against these three antigens all have the effect of inhibiting influenza virus infecti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/11C07K1/04C07K1/06A61K39/145A61P31/16
CPCA61K39/12A61K2039/552A61P31/16C07K14/005C12N2760/16134Y02P20/55
Inventor 肖进宋芳闫福栋胡艳欣巴利民
Owner CHINA ANIMAL HUSBANDRY IND
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