Figure 5: Detection of MASP proteins in ICs by immunocapture assay.

(a) Schematic representation of the methodology followed in the immunocapture assays. (b) Detection of MASP SPs (capture antibody) in EVs from Chagas disease clinical group (target) using anti-EVs Trypo immunoglobulins (detection antibody) (c). Detection of free-IgGs EVs from a pool of each Chagas clinical sera group (target), using anti-SP MAPS as capture antibody and a dilution (1/100) of sera without ICs from Chagas disease clinical group as detection antibody. (d) T.cruzi-specificity of EVs in free-IgGs EVs using anti-TPTC immunoglobulin (T. cruzi positive control) and anti-CD9 antibodies (human positive control) as detection antibodies. (e) Detection of parasite EVs (target) in ICs by immunocapture assays using anti-MASP SP antibodies (captured antibody) and a dilution of a pool of chagasic patients’ sera (1/100) (detection antibody). Turkey- Kramer test multiple comparisons was used. Values of p < 0.001 (***) were considered as significantly different. Cut-off values were obtained calculating the mean of negative controls (sera of non-infected individuals) ±3 (SEM).