Antibody Profiling of Chagas Disease

For mice, ELISA plates were coated with 1 μg/well of rASP-2, rTS or TRASP, reacted with sera diluted 1:200, followed by incubation with secondary antibodies anti-total IgG, anti-IgG1, or anti-IgG2c conjugated with streptavidin-HRP (Cat 1030-05, 1070-05, 1079-05, Southern Biotech), all diluted 1:5000. For dogs, we used ELISA plates (Costar) coated with 100 ng/well of T. cruzi recombinant antigens, sera diluted 1:100 and anti-total IgG, IgG1, IgG2, and IgM conjugated with streptavidin-HRP (Cat A40-123P, A40-120P, A40121P, BEYA40-116P, Bethyl Laboratories, INC), all diluted 1:100,000. The reactions were developed with the substrate 3,3’ 5,5’-tetrametylbenzidine (TMB) from SIGMA and read at 450 nm^{19 (link),73 (link)}.