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Bird flu synthetic peptide vaccine and preparation method thereof

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

Active Publication Date: 2010-01-20
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] 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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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 : Solid Phase Synthesis of Polypeptide Antigens for Synthetic Peptide Vaccines

[0039] 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.

[0040] 1. Preparation of synthetic raw materials

[0041] The synthetic peptide vaccine polypeptide antigen sequence is the amino acid sequence shown in SEQ ID NO.1, the amino acid sequence shown in SEQ ID NO.2 and (SEQ ID NO.1) 2 -K-εK-CONH 2 Polymerized peptides of the indicated amino acid sequence...

Embodiment 2

[0053] Example 2 : Formulation of Synthetic Peptide Vaccines

[0054] This example is the formulation of the synthetic peptide vaccine provided by the present invention.

[0055] 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 it stand for 5 minutes, and then get the synthetic peptide vaccine against avian influenza virus after subpackaging, that is, the synthetic peptide whose amino acid sequence is shown in SEQ ID NO.1 Vac...

Embodiment 3

[0056] Example 3 : Efficacy Test of Synthetic Peptide Vaccines

[0057] In this example, the efficacy test of the synthetic peptide vaccine provided by the present invention is carried out, and the specific pathogen-free (SPF) chicken hemagglutination inhibition test is used to measure the immune effect of the synthetic peptide vaccine.

[0058] The 4-5 week-old SPF chickens were randomly divided into four groups, the first three groups had 10 SPF chickens in each group, and the control group had 5 SPF chickens. The specific grouping situation is shown in Table 1.

[0059] Table 1 Vaccine grouping and number of animals required

[0060] group

[0061] Each SPF chicken neck of the first three groups was subcutaneously injected with vaccine 0.3ml, and the control group SPF chicken neck was subcutaneously injected with normal saline 0.3ml. On the 21st day after inoculation, blood was collected from 5 control chickens, the serum was separated, and the HI antibody tite...

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Abstract

The invention provides a bird flu synthetic peptide vaccine and a preparation method thereof, in particular to polypeptide used for the bird flu synthetic peptide vaccine or a polymer thereof, as well as a vaccine containing the polypeptide or the polymer thereof and both preparation methods thereof, wherein an amino acid sequence of the polypeptide is an amino acid sequence showed as SEQ ID NO.1 or SEQ ID NO.2. The vaccine made from the polypeptide or the polymer thereof can effectively deal to antigenic variation of bird flu virus, has no biological safety problem, can be synthesized easily in large scale and has favorable application prospects.

Description

technical field [0001] The present invention relates to a polypeptide or its polypeptide polymer used for avian influenza synthetic peptide vaccine, a vaccine containing the polypeptide or its polypeptide polymer and their preparation method. Background technique [0002] Avian influenza (Avian influenza, AI) is an infectious disease caused by avian influenza virus (AIV), and it is listed as a class A infectious disease by the International Veterinary Office (OIE) animal diseases. AIV is a type A influenza virus belonging to the Orthomyxoviridae family Influenzavirus. 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 proteins M1 and M2, and nonstructural proteins NS1 and NS2, in which hemagglutinin (HA), neuraminidase (NA) and M2 proteins are three surface antigen molecules of av...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/11A61K39/145A61P31/16C07K1/06C07K1/04
Inventor 齐鹏郭丽清肖进李丙首巴利民宋芳
Owner CHINA ANIMAL HUSBANDRY IND
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