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Polypeptide for resisting novel coronavirus and application thereof
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A technology of viruses and peptides, applied in the application field of drugs or vaccines, can solve the problems of reducing the protective effect
Active Publication Date: 2022-05-06
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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At present, although a variety of vaccines and monoclonalantibody drugs against SARS-CoV-2 have been approved for emergency use, most studies have shown that they are effective against the currently widespread variant strains of the new coronavirus, such as Alpha (B.1.1.7) , Beta (B.1.351) and Gamma (P.1), etc., showed a reduced protective effect, so it is urgent to develop broad-spectrum drugs or vaccines against the current epidemic and new mutant strains that will appear in the future
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[0076] First, hACE2 gene and flag tag were fused and expressed, and the sequence was artificially synthesized (synthesis service provided by Suzhou Jinweizhi), and then connected to the pLVX lentiviral expression vector to obtain the hACE2 expression vector pLVX-hACE2.
[0078] a. Cell preparation: HEK293T cells were plated in 6-well cellculture plates one day in advance, so that the cell confluency density reached 80%-90% the next day.
[0079] b. Transfection: Mix the target plasmids (pLVX-hACE2 and lentiviral packaging plasmids psPAX2 and pMD2.G) at a ratio of 2:2:1, and then transfect after mixing according to the ratio of plasmids and PEI at 1:3. After 4-6 hours, the cell culture medium was replaced with DMEM medium containing 2% FBS, and the culture was continued for 48 hours.
[0080] c. Virus collection and infection: the cell culture supernatant, which contains the packag...
Embodiment 2
[0084] Packaging of SARS-CoV-2 original strain and variant strain pseudovirus
[0085] Preparation of expression plasmid of truncated S protein
[0086] 1) The coding SARS-CoV-2 original strain (WT) and mutant strains (D614G, Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Kappa (B.1.617 .1) and Delta (B.1.617.2)) the last 18 amino acid nucleotides of the S protein are removed to obtain the nucleotide sequences SARS-CoV-2-WT-S-del18, D614G-S-del18 respectively , B.1.1.7-S-del18, B.1.351-S-del18, P.1-S-del18, B.1.617.1-S-del18, B.1.617.2-S-del18 (the sequences are respectively shown in SEQ ID NO:13-19), and was synthesized by Suzhou Jinweizhi Company.
[0087] 2) Cloning each nucleotide sequence obtained in 1) into the pCAGGS expression vector to obtain expression plasmids pCAGGS-SARS-CoV-2-WT-S-del18, pCAGGS-D614G-S-del18, pCAGGS-B. 1.1.7-S-del18, pCAGGS-B.1.351-S-del18, pCAGGS-P.1-S-del18, pCAGGS-B.1.617.1-S-del18, pCAGGS-B.1.617.2-S- del18.
[0088] Packaging of SARS-...
Embodiment 3
[0095] Evaluation of the inhibitory effect of peptides on the original strain of SARS-CoV-2 and its variant strains of pseudoviruses
[0096] This embodiment aims to determine the effect of polypeptide P3 and its five derived polypeptides (P3-1, P3-2, P3-3, P3-4 and P3-5) on the original strain (SARS-CoV-2WT) and variant strains (D614G, Alpha(B.1.1.7), Beta(B.1.351), Gamma(P.1), Kappa(B.1.617.1) and Delta(B.1.617.2)) pseudovirus inhibitory effect .
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Abstract
The invention relates to a polypeptide for preventing or treating new coronavirus and application thereof, the polypeptide is P3 polypeptide and at least one of P3-1 polypeptide, P3-2 polypeptide, P3-3 polypeptide, P3-4 polypeptide and P3-5 polypeptide derived from the P3 polypeptide, and the amino acid sequences of the polypeptide are respectively shown as SEQ ID NO: 1-6. The polypeptide disclosed by the invention has a relatively strong inhibition effect on original strains and various variant strains of the new coronavirus, and can be used for preparing medicines or vaccines for preventing and / or treating diseases caused by the new coronavirus, and the polypeptide is expected to have potential of preventing and / or treating new variant strains and arbecovirus appearing in the future.
Description
technical field [0001] The present invention relates to the field of biomedicine, in particular to polypeptides for preventing or treating new coronaviruses, polynucleotides encoding them, nucleic acid constructs comprising the polynucleotides, expression vectors comprising the nucleic acid constructs, transformed cells, and The above-mentioned pharmaceutical compositions, and their application in the preparation of drugs or vaccines for preventing and / or treating the new coronavirus. Background technique [0002] The SARS-CoV-2 virus is the pathogen of the new coronary pneumonia epidemic, and it is the third coronavirus that has caused human epidemics since the 21st century. At present, although a variety of vaccines and monoclonalantibody drugs against SARS-CoV-2 have been approved for emergency use, most studies have shown that they are effective against the currently widespread variant strains of the new coronavirus, such as Alpha (B.1.1.7) , Beta (B.1.351) and Gamma (...
Claims
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Application Information
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