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Synthetic polypeptide and application thereof, and anti-influenza-virus vaccine

An anti-influenza virus and synthetic peptide technology, applied in the field of immunology, can solve the problems of heavy workload, no exposure, and inability to guarantee 100% consistency, etc., and achieve the effect of simple design, low cost, and high conservatism

Active Publication Date: 2015-12-30
CHANGCHUN BCHT BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problem of active immunization, some researchers simulated the three-dimensional conformation of the whole protein of HA2 to prepare the vaccine as the immunogen. The resulting vaccine is not immunogenic
At the same time, the large workload and high cost are also a major drawback of this method.
Some researchers also consider using relatively cheap and simple whole viruses for vaccine preparation, but HA2 is not exposed to natural viruses, antibodies induced by using whole viruses as immunogens cannot recognize HA2, and neutralization against HA2 cannot be induced by whole viruses Antibody
In addition, the whole virus immunization method is extremely complicated, and the cost is relatively high compared with synthetic peptide vaccines

Method used

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  • Synthetic polypeptide and application thereof, and anti-influenza-virus vaccine
  • Synthetic polypeptide and application thereof, and anti-influenza-virus vaccine
  • Synthetic polypeptide and application thereof, and anti-influenza-virus vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Epitope and conservation detection

[0035] Sequence conservation was evaluated by sequence alignment analysis, and the sequence alignment database consisted of HA2 sequences of 10,763 strains of H3N2 subtype viruses from 1968 to 2014. The application software is ClustalWv1.4 and MEGA5.0, and the HA2 sequences of 1, 2, 3, 5, 7, and B subtype viruses are combined for conservative evaluation. The comparison results show that the synthetic polypeptide of the present invention has a certain degree of conservation with HA2 of subtypes 1, 2, 3, 5 and 7, and the conservation is as high as 97%.

[0036] At the same time, through the comprehensive analysis of vaccine accessibility, flexibility, and hydrophilicity, B / T cell epitopes were detected on the IEDB website, and the results showed that the synthetic polypeptide of the present invention also has B / T cell epitopes.

Embodiment 2

[0037] Example 2: Binding activity and antibody typing of synthetic polypeptide post-immunization serum to influenza virus strain HA protein

[0038] Animal immunization: BALB / c female mice aged 4-6 weeks were immunized, the immunization period was two weeks, and the immunogen was P1-P6 coupled KLH carrier. The immunization dose was 20ug / rat, and the immune adjuvant was Freund's adjuvant (Beijing Dingguo) mixed in a volume of 1:1, and blood was collected before each immunization. Four immunizations were performed.

[0039] A / Brisbane / 10 / 2007 (H3) HA protein was mixed with coating solution (final concentration 5-10 μg / mL) and coated in a 96-well plate, overnight at 4°C, blocked with 1% BSA / PBS at 37°C for 1 hour. Washed with PBST for 3 times, the serum after the first immunization was used as the primary antibody, diluted in PBST, the first dilution was 1:200, 5-fold serial dilution, and incubated at 37°C for 1.5h. Wash 3 times with PBST, add goat anti-mouse typing antibody t...

Embodiment 3

[0041] Example 3: Binding activity of synthetic polypeptide post-immunization serum to HA protein of different subtype influenza virus strains

[0042] Animal immunization: BALB / c female mice aged 4-6 weeks were immunized, the immunization period was two weeks, and the immunogen was P1-P6 coupled KLH carrier. The immunization dose was 20ug / rat, and the immune adjuvant was Freund's adjuvant (Beijing Dingguo) mixed in a volume of 1:1, and blood was collected before each immunization. Four immunizations were performed.

[0043] A / Brisbane / 10 / 2007(H3) HA protein, A / California / 04 / 2009(H1N1) HA protein, A / Canada / 720 / 2005(H2N2) HA protein, A / Anhui / 1 / 2005(H5N1) ) HA protein, A / Netherlands / 219 / 03 (H7N7) HA protein mixed with coating solution (final concentration 5-10μg / mL) coated in 96-well plate, overnight at 4°C, 1% BSA / PBS at 37°C Closed for 1h. Washed with PBST for 3 times, the serum after the first immunization was used as the primary antibody, diluted in PBST, the first diluti...

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Abstract

The invention relates to the technical field of immunology, and discloses a synthetic polypeptide and application thereof, and an anti-influenza-virus vaccine. The amino acid sequence of the synthetic polypeptide is disclosed as SEQ ID NO:1. A novel immunogenic synthetic polypeptide only comprising 15 amino acids is designed according to the neck region in the HA2 antigen region. The synthetic polypeptide has T / B cell epitopes, and can induce humoral immunity. The synthetic polypeptide combines multiple influenza virus subtypes H1, H2, H3, H5 and H7, and has neutralization activity for various subtype H3 strains. Compared with other synthetic polypeptides for the antigen region, the synthetic polypeptide disclosed by the invention has the maximum activity, has the characteristics of high titer, high conservative property and broad spectrum, and has huge advantages as various influenza virus vaccine subtypes.

Description

technical field [0001] The invention relates to the technical field of immunology, in particular to a synthetic polypeptide and its application and an anti-influenza virus vaccine. Background technique [0002] The outbreak of influenza virus is one of the epidemic diseases that have the greatest impact on human production and life so far. The three major influenza epidemics that broke out in the last century were all caused by influenza viruses, causing tens of millions of deaths and causing huge harm and losses to human health and the economy. According to WHO statistics, about 1 billion people in the world are infected with influenza virus every year, and 500,000 to 1,000,000 people die from influenza. The division of different influenza virus subtypes is based on the sequences of hemagglutinin HA and neuraminidase. The main influenza virus subtypes circulating in the population at this stage are A H1N1, H3N2 and B influenza viruses. [0003] The most effective way to p...

Claims

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Application Information

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IPC IPC(8): C07K14/11A61K39/145A61P31/16
CPCA61K2039/545C07K14/005C12N2760/16122C12N2760/16134
Inventor 单亚明孔维姜春来龚鑫石玉华尹贺
Owner CHANGCHUN BCHT BIOTECH
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