Polypeptide or derivative thereof and application of polypeptide or derivative in influenza virus infection
A technology of influenza virus infection and peptide derivatives, applied in the field of novel polypeptide or protein drugs
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Embodiment 1
[0038] Example 1: Chemical Synthesis of Influenza Virus Hemagglutinin Peptide
[0039] According to the amino acid sequence of SEQ ID NO: 1-8 in the sequence listing, a peptide molecule (18 amino acids in length) with a purity greater than 90% was synthesized by using the solid-phase peptide synthesis method (SYMPHONY type 12-channel peptide synthesizer), and passed through HPLC liquid Purity and molecular weight were identified by phase chromatography and MS mass spectrometry. The HPLC purity identification chart and mass spectrogram of SEQ ID NO: 1-8 are shown in Figure 2.
Embodiment 2
[0040] Example 2: Preparation of highly pathogenic H5N1 influenza recombinant virus and determination of the inhibitory activity of influenza virus hemagglutinin peptide on highly pathogenic H5N1 influenza recombinant virus infection
[0041] 1) Preparation of highly pathogenic H5N1 influenza recombinant virus
[0042] The preparation method of the highly pathogenic H5N1 influenza virus is as follows: the HIV vector plasmid (pNL4-3.Luc.R.E., obtained from NIH) and the plasmid of HA cloned into the mammalian cell expression vector pcDNA3.1 are co-transfected into 293T cells ( Human embryonic kidney cells) to produce recombinant virus particles with HIV as the core and wrapped with HA coat protein. The virus particle has the following characteristics: 1) Since the coat protein is hemagglutinin, the selectivity of the virus particle to the host cell depends on the characteristics of the hemagglutinin. The HA protein in the experiment is derived from the highly pathogenic H5N1 in...
Embodiment 3
[0047] Example 3: Preparation of Seasonal H1N1 Human Influenza Virus and Determination of Influenza Virus Hemagglutinin Peptides Inhibiting Pathogenicity of Host Cells Infected by Seasonal H1N1 Human Influenza Virus
[0048] 1) Preparation of seasonal H1N1 human influenza virus:
[0049] In order to detect the effect of influenza virus hemagglutinin peptides on different species and strains of influenza virus infection, we prepared seasonal H1N1 human influenza virus A / Puerto Rico / 8 / 1934 (H1N1) strains that infect humans, and the preparation method The method is as follows: inoculate the allantoic cavity and amniotic cavity of 9-day-old chicken embryos with the virus stock solution, culture the chicken embryos at 37°C for 2-3 days, harvest the virus in the allantoic fluid and amniotic fluid, centrifuge, aliquot, and store at -70°C. Select MDCK cells (canine kidney cells), a sensitive cell line suitable for the growth of influenza virus, as virus-infected cells, DMEM+0.2%BSA+2μ...
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