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Avian influenza virus head protein as well as preparation method and application thereof

A bird flu virus and protein technology, applied in biochemical equipment and methods, virus peptides, antiviral agents, etc., can solve the problems of long production cycle and high production cost, reduce production cost, shorten production time, and ensure biological safety Effect

Active Publication Date: 2013-04-17
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It faces the problem of long production cycle and high production cost in the production process, and needs a large amount of SPF chicken embryos in the production process

Method used

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  • Avian influenza virus head protein as well as preparation method and application thereof
  • Avian influenza virus head protein as well as preparation method and application thereof
  • Avian influenza virus head protein as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Influenza virus protein

[0029] The amino acid composition of the influenza protein is shown in SEQ ID NO.1.

[0030] SEQ ID NO.1:PLILRDCSVAGWLLGNPMCDEFLNVPEWSYIVEKINPANDLCYPGNFNDYEELKHLLSRINHFEKIQIIPKSSWSDHEASSGVSSACPYQGRSSFFRNVVWLIKKNNAYPTIKRSYNNTNQEDLLVLWGIHHPNDAAEQTRLYQNPTTYISVGTSTLNQRLVPKIATRSKVNGQSGRMEFFWTILKPNDAINFESNGNFIAPENAYKIVKKGD

[0031] The amino acid composition of the influenza virus protein can also be: a protein whose amino acid sequence in the protein shown in SEQ ID NO.1 is substituted, deleted or added with one or several amino acids and has influenza protein antigenicity, such as SEQ ID NO.2 :

[0032] PLILRDCSVAGWLLGNPMCDEFINVPEWSYIVEKASPANDLCYPGNFNDYEELKHLLSRINHFEKIQIIPKSSWSNHDASSGVSSACPYLGRSSFFRNVVWLIKKNSAYPTIKRSYNNTNQEDLLVLWGIHHPNDAAEQTKLYQNPTTYISVGTSTLNQRLVPEIATRPKVNGQSGRMEFFWTILKPNDAINFESNGNFIAPEYAYKIVKKGD;或SEQ ID NO.3:

[0033] PLILRDCSVAGWLLGNPMCDEFINVPEWSYIVEKANPANDLCYPGNFNDYEELKHLLSRINHFEKIQIIPKSSWSDHEASSGVSSACPYQGVPSFFRNVV...

Embodiment 2

[0034] Example 2 Preparation of influenza virus protein

[0035] 1. Obtaining the target gene

[0036] The influenza virus A / Bar-headed Goose / Qinghai / 60 / 05(H5N1) GenBank: DQ095615.1 selected in this experiment. According to the sequence of GenBank: DQ095615.1, the nucleotide sequence of SEQ ID NO.4 was artificially synthesized (this synthesis was provided by Shanghai Sangon Bioengineering Technology Service Co., Ltd.),

[0037] atgcctctaat tttaagagat tgtagtgtag ctggatggct cctcgggaac ccaatgtgtg acgaattcct caatgtgccg gaatggtctt acatagtgga gaagatcaat ccagccaatg acctctgtta cccagggaat ttcaacgact atgaagaact gaaacaccta ttgagcagaa taaaccattt tgagaaaatt cagatcatcc ccaaaagttc ttggtcagat catgaagcct catcaggggt gagctcagca tgtccatacc agggaaggtc ctcctttttt agaaatgtgg tatggcttat caaaaagaac aatgcatacc caacaataaa gagaagttac aataatacca accaagaaga tcttttggta ctgtggggga ttcaccatcc aaatgatgcg gcagagcaga caaggctcta tcaaaaccca accacctata tttccgttgg gacatcaaca ctaaaccaga gattggtacc aaaaatagct a...

Embodiment 3

[0062] Example 3 Protein Animal Test

[0063] H5HA head region protein immunogenicity experiment:

[0064] Experimental animals: 24 SPF chickens, each group of 6 divided into 4 groups, namely: blank control group, adjuvant control group, 50 μg antigen amount group, 100 μg antigen amount group

[0065] Reagents: complete Freund's adjuvant and incomplete Freund's adjuvant were purchased from Sigma

[0066] Methods: 50 μg and 100 μg of purified H5HA head region recombinant protein were mixed with Freund's complete adjuvant for intramuscular injection immunization, and 3 weeks later, the corresponding antigen was mixed with incomplete Freund's adjuvant for booster immunization. At the same time, a blank control group and an adjuvant control group were set up. Chicken serum was collected before immunization and 10 days after the last immunization. On the 21st day after the last immunization, 50 μL containing 10 6 EID 50 A Bar-headed Goose Qinghai / 1 / 2005 (H5N1) v...

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Abstract

The invention discloses avian influenza virus head protein as well as a preparation method and application thereof. An amino acid sequence of the avian influenza virus head protein is shown as SEQ ID NO.1. By using the method provided by the invention, the yield of influenza virus HA can be improved, and every two liters of culture medium can obtain larger than 200 mg of purified HA protein. A batch of purified protein can be obtained for 5-6 days, and the production time is shortened. In addition, because the influenza virus can not be used, the biological safety of vaccines can be ensured, and the risk of poison spreading can not occur; and chick embryo virus amplification is not needed, therefore the production cost is greatly reduced, and the invention has favorable application prospect.

Description

technical field [0001] The invention relates to an avian influenza virus protein and its preparation method and application, in particular to a highly expressed functional avian influenza head protein. Background technique [0002] Influenza (abbreviated as influenza) (influenza) is a kind of acute infectious disease caused by influenza virus caused by humans, poultry and animals. Influenza viruses belong to the family Orthomyxoviridae. According to the characteristics of viral nucleocapside protein (NP) and matrix protein (matrix protein), influenza viruses are divided into three types: A (A), B (B) and C (C). Influenza A viruses can be further divided into different subtypes according to the antigenicity of the hemagglutinin (HAmagglutinin, HA) and neuraminidase (neuraminidase, NA) proteins on the surface of virus particles. There are 16 HA subtypes and 9 NA subtypes of influenza A virus found so far. Influenza A virus was discovered in 1901. However, human influenza A...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/11C12N1/21C12N15/70A61K39/145A61P11/00A61P31/16C12R1/19
Inventor 高福宣春玲
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI