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H5 subtype avian influenza virus HA protein B cell epitope polypeptide and application thereof

A technology of avian influenza virus and antigenic epitope, applied in the field of molecular immunology, can solve the difficulty of prevention and control of large avian influenza, simultaneous infection of pigs and other problems

Inactive Publication Date: 2016-11-23
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, α-2,6 receptors and α-2,3 receptors are distributed in pigs, so it can cause human influenza and bird flu to infect pigs at the same time, and recombination occurs in pigs to produce new strains. Difficulty in the prevention and control of bird flu

Method used

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  • H5 subtype avian influenza virus HA protein B cell epitope polypeptide and application thereof
  • H5 subtype avian influenza virus HA protein B cell epitope polypeptide and application thereof
  • H5 subtype avian influenza virus HA protein B cell epitope polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1H5

[0020] Example 1 Identification of H5 subtype avian influenza virus HA protein B cell antigen epitope polypeptide

[0021] 1. Materials and Methods

[0022] 1.1 Main experimental materials

[0023] The H5 subtype avian influenza virus chicken serum was prepared by our laboratory; the synthetic H5 subtype avian influenza virus HA gene polypeptide sequence was referenced (A / DuTurkey / England / N28 / 1973); the epitope peptide chip synthesis service was provided by PEPperPRINT GmbH.

[0024] 1.2 Main reagents

[0025] Washing solution: add 0.05% Tween 20 to PBS with pH 7.4; blocking buffer: Rockland blockingbuffer MB-070; incubation buffer: add 0.05% Tween 20 and 10% Rockland blocking buffer to PBS with pH 7.4; goat anti-chicken secondary antibody: Goat anti-chicken IgG-H+L-DyLight680; control antibody 1: HA(12CA5)-LL-DyLight680; control antibody 2: FLAG(M2)-LL-DyLight800.

[0026] 1.3 Preparation of peptide chip matrix

[0027] The screened H5 subtype HA nucleic acid sequence was...

Embodiment 2

[0040] Example 2 Use of the B cell antigen epitope polypeptide of the avian influenza virus HA protein of the present invention in the diagnosis or detection of H5 subtype avian influenza virus

[0041] 1 Materials and methods

[0042] 1.1 Main experimental materials

[0043] BSA-conjugated peptide service was provided by Shanghai Sangon Bioengineering (Shanghai) Co., Ltd.; H1-H15HA single-factor chicken serum was prepared by our laboratory; high-adsorption enzyme plate was purchased from Coster.

[0044] 1.2 Main reagents

[0045] Bovine serum albumin (BSA) was purchased from MP Biomedicals, USA; HRP-labeled goat anti-chicken antibody was purchased from Sigma, USA; TMB chromogenic solution was purchased from ABM, USA.

[0046] 1.3 Synthesis of H5-1, H5-2, H5-3, H5-4 epitope polypeptides

[0047] The four H5 subtype HA antigen epitope polypeptides screened by the HA antigen matrix chip in Example 1 were artificially synthesized, and a flexible amino acid C was added to the ...

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Abstract

The invention discloses H5 subtype avian influenza virus HA protein B cell epitope polypeptide and application thereof. An H5 subtype AIV (avian influenza virus) HA amino acid sequence is used as the template, the epitope polypeptide is synthesized on a solid-phase carrier in situ, each polypeptide contains 15 amino acids, and 14 amino acids of each previous polypeptide and the corresponding next polypeptide overlap. The synthesized H5 AIV HA epitope is allowed to have reaction with known H5 subtype positive serum, and one epitope polypeptide (H5-1) which can have specific reaction with H5 AIV chicken serum. The epitope polypeptide is used for detecting H5 subtype avian influenza viruses, and the detection result shows that the ELISA method built by the epitope polypeptide is suitable for detecting a 1:200 diluted serum sample. A coincidence rate test proves that the detection result of epitope polypeptide on H5 AIV negative serum is negative, the detection coincidence rate of samples with the hemagglutination inhibition values ranging from log2 to log4 is 87.6%, and the detection coincidence rate of samples with the hemagglutination inhibition values being log5 or above log5 is 98.3%.

Description

technical field [0001] The present invention relates to antigenic epitope polypeptides, especially to H5 subtype avian influenza virus HA protein B cell antigenic epitope polypeptides. The present invention also relates to the application of the antigenic epitope polypeptides in the preparation of drugs for diagnosing or detecting avian influenza virus infection, belonging to molecular field of immunology. Background technique [0002] Avian influenza (Avian influenza, AI) is an infectious disease mainly spread in poultry caused by type A avian influenza virus (AIV). AIV can cause respiratory diseases after infecting poultry, resulting in a decline in egg production, and can cause 100% mortality in severe cases. [0003] In recent years, AIV has frequently mutated, making some mutant strains directly infect humans and cause death, seriously endangering public health and causing huge economic losses, so it has been closely watched. On-the-spot detection mainly judges the im...

Claims

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Application Information

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IPC IPC(8): C07K14/11C12N15/44C12N15/63G01N33/68G01N33/569
CPCC07K14/005C12N2760/16122G01N33/56983G01N33/6803G01N2333/11G01N2469/20
Inventor 王秀荣包红梅陈化兰
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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