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. 2023 Dec 17;11(9):2303366. doi: 10.1002/advs.202303366

Figure 2.

Figure 2

Immunization of Mosaic RBD‐np induce robust humoral response in mice. A) Scheme of the immunization process in mice. B) Six‐week‐old female BALB/c mice were immunized with 1, 5, or 10 µg mosaic RBD‐np. Mice were boosted 3 weeks with their respective antigen dose after primary vaccination. Neutralization antibody were detected by focus reduction neutralization test (FRNT) on day 7 and day 21 after boost immunization. n = 2, 3 or 4, Bars indicate median values. C) Six‐week‐old female BALB/c mice were immunized with 10 µg mosaic RBD‐np/SAS. Mice were boosted 3 weeks with their respective antigen dose after primary vaccination. IgG responses in the sera of vaccinated mice were evaluated two weeks after priming or boosting by ELISA for binding to SARS‐CoV‐2 (WT), MERS‐CoV and SARS‐CoV RBDs. n = 3 mice per group, and the serum were pooled by group. Three independent experiments were analysed. D) Neutralization antibody titre of SARS‐CoV were detected by plaque reduction neutralization test (PRNT). Neutralization antibody titre of SARS‐CoV‐2 WT and MERS‐CoV EMC were detected by FRNT. n = 3‐5 mice per group. E) Neutralization antibody titre of SARS‐CoV‐2 VOCs were detected by FRNT. n = 3‐5 mice per group. F) Neutralization titre of MERS‐CoV GD01, Nigeria, and SARS‐rCoV WIV‐1, Pangolin were detected by pseuovirus assay. Statistical analyses were performed using two‐way ANOVA. Data represent two independent experiments. All results are expressed as mean ± SEM. ns, no significantly difference, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.