a fusion PspA protein minus the proline rich domain was poorly expressed in Salmonella and immunization with an RASV revealing this fusion gene offered only weak protection in mice. This suggests that the addition of longer PspA parts, containing more protected regions, not merely is important for broadening cross safety against strains of different PspA clades but additionally improves fusion gene expression in Salmonella. In a recent review, Darrieux et al. Produced two family 1 family 2 fusion proteins and PspA subclones like the proline rich regions and helical regions from family 1 and family 2 strains. The proteins were purified from E. coli. Groups of natural product library mice were injected with three doses of each protein. Serum was evaluated for surface binding and complement deposition. The results because study were similar to our results with regard to family specific responses against clade 1 and clade 4 strains. However, in our study, we observed cross clade binding and complement deposition against clade 3 and 2 strains that were not observed in the last study. This big difference in results might be due to differences in amino acid sequence and fusion protein structure or due to the method of delivery: mucosal immunization with an RASV versus parenteral injection. We observe that their fusion proteins were derived from different S. pneumoniae traces from those used here. Mice immunized with three doses of the purified proteins Chromoblastomycosis were partially protected against challenge with S. pneumoniae strains A66. 1, 679/99, and 3JYP2670. But, neither protein protected well against all three ranges. Our results showed that both fusion proteins offered substantial protection against challenge, irrespective of challenge pressure or challenge course. Additionally, one of the fusion proteins we used, PspA/Rx1 EF5668, elicited a strong immune response, eliciting serum antibodies that bound avidly to strains from all five clades tested, successfully focused complement deposition on these strains, and provided somewhat better security to challenge than the other PspA proteins tested. These results show that the PspA blend protein managed enough architectural epitopes Cathepsin Inhibitor 1 so that antibodies against them could bind to native PspA on intact bacteria. The binding of antibody for the bacteria was in keeping with the ELISA results. Taken together, these results show that PspA combination delivery by RASV may lead to a more generally protective immune response than parenteral injection. It’ll be interesting to find out whether this is established in future studies using identical protein fusions. In Western blots of the RASV traces synthesizing fusion proteins PspA/Rx1 EF5668 and PspA/EF5668 Rx1, we discovered numerous bands smaller than the expected 107 kDa size of the whole protein. There have been small bands reacting with anti PspA/EF5668 antibodies and anti PspA/Rx1, though we could not decide whether any bands besides the size of the total size protein reacted with both antibodies.