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. 2023 Jan 13;26(2):105957. doi: 10.1016/j.isci.2023.105957

Figure 2.

Figure 2

Sera and purified IgG from MPXV recoverees bind recombinantly expressed MPXV antigens

(A) Table listing the three MPXV antigens A35R, M1R and H3L produced in this study. Published PDB IDs of VACV homologs are given.29,30,31 Amino-acid sequence alignment scores between MPXV and VACV antigens were calculated using the Clustal Omega web tool.32

(B) Construct design of the three recombinantly expressed antigens, A35R, M1R and H3L. Signal peptide, His-tag and Avi-tag are in black, light gray and dark gray, respectively. The amino acid section produced for each antigen is stated.

(C) SDS-PAGE (left) and western blot using mouse anti-Avi-tag antibody (right) of the recombinantly expressed MPXV antigens A35R, M1R and H3L after purification. The antigens are indicated on top, the protein marker is on the right of each gel/blot, and protein sizes are indicated on the right in kDa.

(D) Serum binding to MPXV antigens A35R, M1R and H3L as detected by ELISA. MPXV recoverees are in red (n = 11), VPXV are in purple (n = 6), uninfected >45yo are in blue (n = 11) and uninfected <45yo are in gray (n = 11). Upper panel: The bold lines and the shadowed areas on the graph represent the mean serum response and SEM, respectively, of 4 consecutive serum dilutions for every group of donors, starting from 1:75 dilution. Lower panel: AUC values for every individual donor. Statistical analysis was performed using One-way ANOVA. ∗p< 0.05, ∗∗p< 0.01, ∗∗∗p< 0.001, ∗∗∗∗p< 0.0001.

(E) Purified IgG binding to MPXV antigens A35R, M1R and H3L as detected by ELISA. MPXV recoverees are in red (n = 11), VPXV are in purple (n = 6), uninfected <45yo are in gray (n = 2), and VIG is represented by the dashed black line. Eight consecutive IgG dilutions starting from 100 μg/mL were tested (x axis). Upper panel: The bold lines and the shadowed areas on the graph represent the mean IgG response and SEM, respectively, for every group of donors. Lower panel: AUC values for every individual donor. Statistical analysis was performed between MPXV and VPXV groups using Welch’s t test. ∗∗p< 0.01.