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Common antigenic epitope, antibody, identification method and application of various subtype influenza virus ha2 proteins

An influenza virus, common antigen technology, applied in the direction of virus antigen components, antiviral immunoglobulin, virus/phage, etc., can solve the problem of not having broad spectrum, and achieve the effect of improving sensitivity and reducing steps

Active Publication Date: 2021-08-24
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Epitopes identified on the HA1 protein of influenza A viruses tend to be subtype-specific and not broad-spectrum

Method used

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  • Common antigenic epitope, antibody, identification method and application of various subtype influenza virus ha2 proteins
  • Common antigenic epitope, antibody, identification method and application of various subtype influenza virus ha2 proteins
  • Common antigenic epitope, antibody, identification method and application of various subtype influenza virus ha2 proteins

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Embodiment

[0033] 1. Serum Preparation

[0034] Table 1 Background information of 13 strains of influenza virus and the hemagglutination inhibitory titer of their immune sera

[0035]

[0036] Preparation of whole virus immunized chicken serum: IAV was inactivated with formaldehyde, mixed with Tween 80 and white oil to make inactivated oil emulsion vaccine, and immunized 3-week-old SPF chickens with a dose of 0.3 mL / bird. A second immunization was performed 2 weeks later with the same dose. Serum was collected in the second week after the second immunization, and the hemagglutination inhibition (HI) titer was greater than or equal to 6log2 for future use (see Table 1 for details).

[0037] Preparation of peptide-immunized chicken serum: 100 μg of BSA-coupled peptide (synthesized by Shanghai GL Biochem Company) was dissolved in 0.1 mL of normal saline and mixed with an equal amount of Freund’s complete adjuvant (Sigma, USA), and subcutaneously immunized at 21 days old SPF chicken. T...

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Abstract

The invention relates to a common antigenic epitope, antibody, identification method and application of multiple subtype influenza virus HA2 proteins, and the antigenic epitope can be combined with H1, H3, H4, H5, H6, H7, H8, H9 and H10 subtype influenza viruses immune serum combination. Select an H5N1 subtype influenza strain S, design 18 polypeptides with a length of 20 amino acids and overlap each other by 10 amino acids according to the sequence of its HA2 protein, artificially synthesize them, place samples on modified silica gel and mold them into a polypeptide chip, use Screening of universal epitopes with antisera against different subtypes of IAV. The recombinant virus with the mutation of the epitope was constructed, and the binding ability of the epitope to the immune serum was analyzed by Western blot. The results showed that 485‑FYHK CDNECME‑495 on HA2 protein had broad-spectrum binding activity to H1, H3, H4, H5, H6, H7, H8, H9 and H10 subtype IAV antisera. K / D mutations in the recombinant virus resulted in reduced serum binding activity, suggesting that they are key amino acids for serum binding activity. The peptide also contains HLA-DR1-restricted CD4 T-cell epitopes.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a common antigenic epitope, antibody, identification method and application of multiple subtype influenza virus HA2 proteins. Background technique [0002] Type A influenza virus (Influenza Avirus, IAV) consists of 8 segments of single-stranded negative-sense RNA, prone to antigenic drift and antigenic shift. Currently, 18 HA subtypes and 11 NA subtypes have been found, among which avian influenza virus (AIV) includes 16 HA subtypes and 9 NA subtypes. According to genetic evolution analysis, these 18 HA subtypes can be divided into two groups, Group 1 is H1, H2, H5, H6, H8, H9, H11, H12, H13, H16 and H17; Group 2 is H3, H4, H7, H10, H14 and H15. Due to the persistent endemic and seasonal epidemics, the antigenicity of IAV of the same subtype has also changed greatly, and different lineages have gradually been derived. The monitoring and control of IAV is also becoming more and mor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/11G01N33/569G01N33/68C07K16/10A61K39/145A61P31/16
CPCA61K39/12A61K2039/552A61P31/16C07K14/005C07K16/1018C12N2760/16122C12N2760/16134G01N33/56983G01N33/6854G01N2333/11G01N2469/10
Inventor 彭大新王秋霞陈素娟秦涛孙志豪
Owner YANGZHOU UNIV