Vaccine composition for resisting H7 subtype avian influenza virus as well as preparation method and application of vaccine composition

A bird flu virus and vaccine composition technology, applied in the field of biopharmaceuticals, can solve problems such as poor stability and lack of immune effect of commercial whole virus vaccines

Pending Publication Date: 2021-12-24
PU LIKE BIO ENG
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the avian influenza virus-like particle vaccine published in literature research in the prior art can produce better immune response, it still cannot reach the immune effect of commercialized whole virus vaccine, and its stability is relatively poor. Therefore, screening ideal The sequence of avia

Method used

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  • Vaccine composition for resisting H7 subtype avian influenza virus as well as preparation method and application of vaccine composition
  • Vaccine composition for resisting H7 subtype avian influenza virus as well as preparation method and application of vaccine composition
  • Vaccine composition for resisting H7 subtype avian influenza virus as well as preparation method and application of vaccine composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1H7

[0034] Expression of embodiment 1H7 subtype avian influenza virus-like particles

[0035] 1. Carrier modification

[0036] Using the commercially available vector pFastBac I as a template, XhoI, SphI, NheI, MluI, EcoRI and XbaI restriction enzyme sites were sequentially inserted at positions 4413-4414. Add 1 μl of Dpn I enzyme to the PCR product to digest the template plasmid, take 5 μl and transform it into DH5α according to the conventional method, and the successfully transformed plasmid is named pFastBac mut.

[0037] 2. Genetic modification

[0038] The amino acid sequence shown in Seq ID No.1 of the H7N9 avian influenza virus HA gene was modified through artificial deletion, insertion and mutation: LWF 526-528 , G 531 、AIVMG 539-543 missing, I 532-533 、CVV 537-538 、MWACQ 547-548 Insertion, I536L, V545G mutations. The amino acid sequence of the modified H7N9 avian influenza virus HA gene is shown in Seq ID No.2.

[0039] 3. Construction of three expression casset...

Embodiment 2H7

[0050] The preparation of embodiment 2H7 subtype avian influenza virus-like particle vaccine

[0051] The virus-like particles harvested in Example 1 were respectively added to the mineral oil adjuvant to prepare various vaccine compositions, and the specific proportions are shown in Table 1.

[0052] Table 1 Proportion of H7 subtype avian influenza virus-like particle vaccine composition

[0053] components Vaccine 1 Vaccine 2 Vaccine 3 rBac-MNH-1 (HA content) 6log2 8log2 - rBac-MNH-2 (HA content) - - 8log2 Adjuvant (V / V%) 66％ 66％ 66％

Embodiment 3H7

[0054] Embodiment 3H7 subtype avian influenza virus-like particle vaccine immunogenicity test

[0055] Get 50 SPF chickens at the age of 21 days and divide them into 5 groups, 10 in each group. The first group to the third group are respectively injected with the vaccines 1 to 3 prepared in Example 2 subcutaneously in the neck, and the fourth group is injected subcutaneously with commercial products. Chemical inactivated vaccine (H5 (Re-11+Re-12), H7 (H7-Re2) trivalent inactivated vaccine, H7HA content is 8log2), immunization dose is 0.3ml, the 5th group injects 0.3ml normal saline subcutaneously, as a blank control. All experimental chickens were kept in isolation, blood was collected on the 14th and 21st day after immunization, serum was separated, and the titer of HI antibody was determined*. The test results of different HI antibodies after immunization are shown in Table 2.

[0056] Table 2 Results of immunogenicity test of H7 subtype avian influenza virus-like particle...

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Abstract

The invention relates to a vaccine composition for resisting H7 subtype avian influenza virus, the vaccine composition comprises an immune dose of H7 subtype avian influenza virus-like particle antigen and a pharmaceutically acceptable carrier, and the H7 subtype avian influenza virus-like particle antigen is assembled by H7 subtype avian influenza virus HA, NA and M1 antigen proteins. The vaccine composition for resisting the H7 subtype avian influenza virus can achieve the immune effect of an inactivated vaccine with a higher immune dose when the immune dose is low, the high-titer antibody is generated in a shorter time, the poultry are protected, the vaccine composition is good in stability, and the stability and durability of immunogenicity of the vaccine composition are advantageously guaranteed.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a virus-like particle vaccine providing protection against avian influenza virus, its preparation method and application. Background technique [0002] Virus-like particles (VLPs) are hollow particles between 15nm and 400nm assembled by structural proteins of viruses. VLPs can be prepared by highly expressing one (or several) structural proteins of a certain virus in vitro, so that they can automatically assemble into hollow particles similar in shape to natural viruses. The method is mainly to clone viral structural protein genes into expression vectors, and then transfer these vectors into prokaryotic or eukaryotic cells for expression. [0003] Avian Influenza Virus (AIV) belongs to the family Orthomyxoviridae, the genus Influenzavirus, and is a type A influenza virus. Avian influenza (Avian Influenza, AI) is a bird infection and disease syndrome caused by the virus. The I...

Claims

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Application Information

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IPC IPC(8): A61K39/145A61P31/16
CPCA61K39/12A61P31/16C12N2760/16134A61K2039/5258A61K2039/552
Inventor 田克恭王同燕张盼涛张许科
Owner PU LIKE BIO ENG
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