An avian influenza vaccine composition, a preparing method thereof and applications of the composition

A vaccine composition and bird flu technology, applied in biochemical equipment and methods, vaccines, veterinary vaccines, etc., can solve problems such as high production costs, waste of manpower and material resources, and long vaccine production cycle, and achieve enhanced delivery , increase the penetrating ability, increase the effect of expression

Inactive Publication Date: 2015-01-21
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the production of avian influenza vaccines is constantly updated with the mutation of the virus, the production cycle of the corresponding vaccine against influenza virus epidemi...

Method used

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  • An avian influenza vaccine composition, a preparing method thereof and applications of the composition
  • An avian influenza vaccine composition, a preparing method thereof and applications of the composition
  • An avian influenza vaccine composition, a preparing method thereof and applications of the composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1 artificial synthesis M2, 3M2e protein

[0046] According to NCBI ( http: / / www.ncbi.nlm.nih.gov ) nucleotide sequences of M2 and M2e protein genes reported in A / Duck / Hong Kong / Y439 / 97 (H9N2, accession number: AF156462) (see sequence table SEQ ID No.1 and SEQ ID No.3 for details), The amino acid polypeptide sequences of M2 and 3M2e proteins were artificially synthesized (see sequence table SEQ ID No.2 and SEQ ID No.4 for details), and the synthesized polypeptide sequences were purified by high performance liquid chromatography to obtain purified M2 and 3M2e proteins respectively . Then, the M2 and 3M2e proteins were respectively prepared into 1 mg / mL stock solution with physiological saline containing 1% trehalose.

Embodiment 2

[0047] The preparation of embodiment 2 influenza vaccines

[0048] The film method of the preparation method of influenza vaccine of the present invention combines with freeze-drying method, comprises the following operation steps:

[0049] (1) Soybean lecithin m1, cholesterol m2, and vitamin E (Ve) m3 were mixed and dissolved in V1 chloroform according to the amount in Table 1, and the solution was placed in an eggplant-shaped bottle of a rotary evaporator, and a water bath at 45°C was used as Evaporation temperature, vacuum rotary evaporation, film formation.

[0050] (2) Add physiological saline V2 according to the final concentration of the film material at 2% (W / W), add a few small glass beads, and rotate to evaporate at 45°C. After the film completely falls off the bottle wall, let it stand for about 25 minutes to make the bottle Medium liquid is milky white suspension.

[0051] (3) Add different concentrations and volumes of avian influenza antigen solution (containin...

Embodiment 3

[0056] Embodiment 3 animal experiments

[0057] A total of 120 3-week-old SPF chickens were selected and randomly divided into 6 groups, 20 birds per group. They were subcutaneously injected with homemade influenza vaccine, blank liposomes, and normal saline (as a blank control) at a dose of 0.3 mL / feather.

[0058] On the 0th day, 15th day, 30th day, 45th day, 60th day, and 90th day after immunization, the serum of each test group was collected and separated. At the same time, the serum anti-M2 or M2e antibody titer was detected by indirect ELISA method. The specific operation steps were as follows: Dilute the M2 or M2e protein stock solution to 1 μg / mL with normal saline, coat the 96-well plate, 50 μL / well, and wrap at 4 °C. overnight; use blocking solution containing 5% skimmed milk powder for blocking; after washing, add the diluted serum to be tested, 50 μL / well, and react at 37°C for 2 hours; after washing, add enzyme-labeled secondary antibody, 50 μL / well, and incubate ...

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Abstract

The invention provides an avian influenza vaccine composition. The composition comprises a liposome, and an immunizing dose of M2 and/or 3M2e protein. The sequence of the M2 protein is SEQ ID NO.2. The sequence of the 3M2e protein is SEQ ID NO.4. According to the avian influenza vaccine, the M2 protein and/or the 3M2e protein which are the main components in an antigen in an influenza vaccine are included by utilization of vesicular microspheres formed by biological membrane bilayers of the liposome. Under functions of the liposome, the vaccine can efficiently carry the antigen included to an antigen presenting cell in a targeted manner, stimulates a body to generate humoral immunity and cellular immunity, has a broad spectrum and durability, resists a plurality of types or subtypes of avian influenza viruses, has a function of slowly releasing the antigen under the actions of the liposome, and is capable of being degraded by the body and free of side and toxic effects. The vaccine is beneficial to large-scale production.

Description

technical field [0001] The invention relates to veterinary biological products, in particular to an avian influenza vaccine and its application in medicines for preventing and / or treating different subtypes of avian influenza virus. Background technique [0002] Avian influenza (Avian influenza, AI) is the abbreviation of avian influenza, the acute respiratory infectious disease that is infected by the multiple poultry that is caused by avian influenza virus (Avian influenza virus, AIV), also can infect human being, is by International Veterinary Bureau (OIE ) and my country's "Livestock and Poultry Epidemic Prevention Regulations" are listed as Class A severe infectious diseases. [0003] Influenza viruses belong to the Orthomyxoviridae family Influenzavirus genus, which contain enveloped, polymorphic negative-strand RNA viruses. Influenza viruses can be divided into three types: A (A), B (B) and C (C) according to the differences in the antigens of the nucleoprotein (NP) a...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61K39/39A61K9/127A61P31/16
CPCA61K9/127A61K39/12A61K39/39A61K2039/552A61K2039/55555C12N2760/16034
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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