Fully human monoclonal antibodies against Zika virus and application thereof
A monoclonal antibody and Zika virus technology, applied in the fields of application, antibody, antiviral agent, etc., can solve the problems of no specific therapeutic drugs and no Zika virus vaccine developed
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[0061] 1) Production of Zika virus E protein
[0062] The synthetic Zika virus E protein gene was inserted into the expression plasmid, and the expression plasmid was transiently transfected into Expi293 cells, and the E protein-human Fc fusion protein was expressed and purified;
[0063] 2) Screening of Zika virus-specific antigen-binding fragments (Fab)
[0064] Using a large phage display antibody library constructed from the cDNA of peripheral blood mononuclear cells from 40 healthy volunteers, antibodies were screened against biotinylated protein E; biotinylated protein E was immobilized on streptavidin-coated magnetic beads up, 10 12 Each phage-displayed Fab was incubated with 5, 4, 2, and 1 micrograms of antigen for two hours in rounds 1, 2, 3, and 4 at room temperature, and 10 phages were used for each round of screening. 12 1; use polyclonal phage ELISA to detect the enrichment of antibodies, the 1st, 2nd, 3rd, and 4th rounds of phages are incubated with coated prot...
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