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Structure and application of influenza virus hemagglutinin protein binding polypeptide

A technology of hemagglutinin protein and influenza virus, which is applied in the field of anti-influenza virus, can solve the problems of influenza virus drug resistance, etc., and achieve the effect of inhibiting infection, inhibiting cytopathic, and inhibiting proliferation

Inactive Publication Date: 2011-12-07
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the widespread application of NA inhibitors in the population, the influenza virus has also developed a certain degree of drug resistance.

Method used

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  • Structure and application of influenza virus hemagglutinin protein binding polypeptide
  • Structure and application of influenza virus hemagglutinin protein binding polypeptide
  • Structure and application of influenza virus hemagglutinin protein binding polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Screening of hemagglutinin protein-binding polypeptides

[0037] Materials and Methods

[0038] 1. Induced expression and purification of GST-HA fusion protein

[0039] pGEX-6p-1-HA was constructed in our laboratory, and the CDS region of HA was amplified from the A / Jingfang / 86-1(H1N1) virus preserved in our laboratory, and inserted into the pGEX-6p-1 plasmid by cloning , the results of positive clone screening, sequencing, and BLAST comparison showed that the constructed pGEX-6p-1-HA was completely correct. Inoculate Escherichia coli DH5α transformed with the vector pGEX-6p-1-HA expressing GST fusion protein into LB medium, shake overnight at 37°C, then inoculate into fresh LB medium at 2% inoculum size, and culture at 30°C After 5 hours, add IPTG to a final concentration of 0.1 mmol / L-1 mmol / L, and continue to induce culture at 16°C or 4°C overnight. After the inclusion bodies expressing the GST-HA fusion protein were dissolved, they were sequentially pur...

Embodiment 2

[0052] Example 2 Anti-influenza Virus Cytopathic Activity of Hemagglutinin Protein Binding Polypeptide

[0053] Materials and Methods

[0054] 1. A549 cells, viruses and control drugs

[0055] The virus was passaged in 10-day-old SPF chicken embryos, the allantoic fluid was collected, and the hemagglutination (Hemagglutination, HA) titer was measured, and the HA price was taken as > 1:2 6 For those, mix well, aliquot and store at -70°C for later use. The virus strain used was A / Jingfang / 86-1 (H1N1). A549 cell culture medium is F12K culture medium containing 10% fetal bovine serum (Gibco), and a maintenance solution containing 0.25% fetal bovine serum and 5 mg / ml trypsin is used for the determination of virus infection and cytopathic changes. The positive control drug was rimantadine.

[0056] 2. Dose-dependent effect of H17 polypeptide on inhibiting cytopathic effect of influenza virus

[0057] The A549 cells were plated on a 96-well cell culture plate, and 60-80% of them...

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Abstract

Belonging to the technical field of biomedicine, the invention relates to a sequence and structure of a polypeptide able to specifically bind with influenza virus hemagglutinin protein, and application of the polypeptide in anti-influenza viruses. By expressing purified influenza virus hemagglutinin protein and screening a random peptide library with a phage display technology, a polypeptide specifically bound with influenza virus hemagglutinin and equipped with sequences numbered 1-18 can be obtained. As a hemagglutinin-binding peptide can hinder the combination of hemagglutinin and a host cell receptor, so the influenza virus can be inhibited from infecting the host cell. Thus, the invention also conducts an anti-influenza virus activity study to the hemagglutinin-binding peptide selected from the phase peptide library, and finds that a polypeptide H17, with a sequence of NH2-SHGRITFAYFAN-COOH, can effectively inhibit the influenza virus from infecting the host cell and is of small toxicity. Therefore, the hemagglutinin-binding peptide of the invention and the H17 polypeptide therein with an anti-influenza virus activity are expected to become novel treatment medicaments for treating diseases caused by influenza virus infection and reducing the hazards of diseases caused by influenza viruses.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and more specifically relates to the sequence and structure of a polypeptide specifically combined with influenza virus (such as H1N1 subtype) and its use in anti-influenza virus. Background technique [0002] Influenza causes about 500 million people to become sick and 250,000 to 500,000 to die every year in the world, and the annual incidence of influenza in my country is 10% to 30%. What is even more worrying is that in recent years, the number of human infections with animal-derived highly pathogenic and highly lethal avian influenza viruses (H5N1) and swine influenza (H1N1) has been increasing, and the mortality rate is quite high. One of the reasons for the raging influenza virus is its strong ability to mutate antigens, which limits the immune protection effect of the vaccine. Moreover, the vaccine can only prevent known influenza virus subtypes, and has no effect on those caused by an...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K7/08A61K38/08A61K38/10A61P31/16
Inventor 王升启丁晓然张莉吴小桃杨静王学军
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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