Fully human monoclonal antibody against h7n9 subtype avian influenza virus and its application
A monoclonal antibody and avian influenza virus technology, applied in the direction of antibodies, applications, antiviral agents, etc., can solve problems such as the prevalence of mutant strains, antigenic variation of influenza viruses, and no therapeutic drugs
Active Publication Date: 2021-05-04
FUDAN UNIV
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Problems solved by technology
The HA antigen variation caused by HA gene mutation often leads to immune failure and the prevalence of mutant strains. Influenza viruses frequently and continuously undergo antigenic variation. At present, there is no effective vaccine against new avian influenza viruses such as H7N9, and there is no specific effect. Therapeutic drugs
Method used
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Embodiment 1
[0057] 1) Preparation of H7N9 subtype avian influenza virus HA protein
[0058] The synthetic H7N9 subtype avian influenza virus HA gene (SH2) was fused with IgG1Fc, inserted into the expression plasmid, and the expression plasmid was transiently transfected into 293FREESTYLE cells to express and purify the HA1(SH2)-Fc protein; Methods express HA1 protein without IgG1 Fc tag, and HA1 protein in trimeric form;
[0059] 2) Screen H7N9 subtype avian influenza virus-specific antigen-binding fragment (Fab)
[0060] Using a large phage display antibody library constructed from the cDNA of peripheral blood mononuclear cells from 40 healthy volunteers, the antibodies were screened against biotin-labeled HA1(SH2)-Fc protein and commercial HA-SH1 and HA-SH2 proteins; 10 12 Each phage-displayed Fab was incubated with 5, 3, 3, and 1 micrograms of antigen for two hours at room temperature in rounds 1, 2, 3, and 4, respectively, and coated with HA1(SH2)-Fc, HA-SH1, and HA-SH2 proteins On...
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The invention belongs to the field of biotechnology, and specifically relates to a fully human monoclonal antibody against H7N9 subtype avian influenza virus, an antigen-binding fragment thereof, and applications thereof. The antibody is determined by the specific gene sequences of the complementarity determining regions (CDRs) present in the variable regions of the antibody light chain and heavy chain genes, and obtains efficient expression in prokaryotic and eukaryotic cells specifically binding to the H7N9 subtype Antibodies to the hemagglutinin protein (HA) of the avian influenza virus. Using the antibody CDR region or part or whole gene, different forms of genetically engineered antibodies can be modified and produced in prokaryotic and eukaryotic cells and any expression system, and can be used to prepare preparations for preventing or treating H7N9 subtype avian influenza virus-related diseases.
Description
Technical field: [0001] The invention belongs to the field of biotechnology, and specifically relates to a fully human monoclonal antibody against H7N9 subtype avian influenza virus, an antigen-binding fragment thereof, and applications thereof. Background technique: [0002] Avian influenza viruses are reported to be influenza viruses that infect birds and do not usually infect humans. However, in 1997, Hong Kong, my country reported the first case of human infection with avian influenza. This highly pathogenic avian influenza virus is of the H5N1 subtype. As of December 10, 2013, a total of 648 cases have been reported in 15 countries and regions around the world, including 384 deaths. Subsequently, multiple subtypes of avian influenza viruses such as H9N2 were also found to have the ability to infect humans. In particular, in March 2013, the first cases of human infection with the new H7N9 subtype avian influenza virus were found in Shanghai and Anhui, my country. At p...
Claims
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IPC IPC(8): C07K16/10C07K16/46C12N15/13C12N15/85C12N5/10G01N33/68G01N33/577G01N33/569A61K39/42A61P31/16
Inventor 应天雷姜世勃陆路贺彪孙志武
Owner FUDAN UNIV