IgG responses to recombinant VM protein immunization. (A) Antibody titers were measured in each study group pre-and post-challenge against individual VM proteins in triplicate using ELISA. Each data line represents the average IgG response of each animal (n-10). Box and whiskers plots represent the antibody titers against t3490, LA3490, LA0620, LA1402, LA1400, and LA0591, respectively. The four-study groups include G-I: PBS, G-II: t3490, G-III: VM mix and G-IV: VM unlabeled. The box boundaries indicate the median and interquartile ranges and the whiskers denote the maximum and minimum values. Statistical analysis was performed by t-test and the non-parametric, unpaired, two tailed Mann Whitney test p < 0.0001 values were considered significant. (B) Aliquot from immunized recombinant purified VM proteins were run in 4 ± 12% SDS-PAGE then transferred to nitrocellulose membrane for Western blot analysis. The membrane was probed probed with pooled sera (1:100 dilution) collected post-challenge. PIB denotes pre-immunized bleed, served as control. VM proteins were recognized by sera from G-II, G-III, and G-IV. Lane 1 shows VM mix protein (LA3490, LA0620, LA1402, LA1400 and LA0591) and Lane 2 shows VM unlabeled proteins (LA1400 and LA0591). Arrows shows expected size of VM proteins. M represents molecular weight markers. **p < 0.05, ***p < 0.001, ****p < 0.0001.