Mouse anti flavivirus 4g2 monoclonal antibody mab

ADE potentially induced by immunized mouse serum antibodies was measured in K562 cells using a flow cytometry-based assay (de Alwis et al., 2014 (link)). Briefly, 100 PFU of ZIKV (strain R103451) was mixed with sera at 4-fold serial dilutions, and incubated at 37°C for 1 h. The virus-serum mixture was then added to K562 cells (5 × 10⁴/well), and incubated at 37°C for 2 h in DMEM containing 10% FBS and 1% Penicillin/Streptomycin, followed by washing the cells with fresh DMEM, and culturing them for 3 days. The cells were then fixed, permeabilized, and sequentially stained with mouse anti-flavivirus 4G2 monoclonal antibody (mAb, 2 μg/ml) and FITC-conjugated anti-mouse antibody (1:100, Biolegend). The percent of infected cells was calculated based on the fluorescence signals in the presence or absence of serially diluted mouse sera.