ELISA Binding Kinetics Characterization

ELISAs were performed as previously described (Hastie et al., 2017 (link), 2019 (link)). Briefly, ELISA plates (Corning, Kennebunk, ME) were coated with 2.5 μg/mL streptavidin in PBS for 3 h at RT, and blocked with 3% BSA in PBS for 2 h at RT. LASV pfGP-TD (1 μg/mL) either alone or in complex with mAb (10 μg/mL) was diluted in PBS for 1hr at RT before coating on ELISA plates (Corning, Kennebunk, ME). After blocking, a dilution series of LAMP1-RbFc starting at 10 μg/mL was bound to coated protein for 2 h at RT in 50 mM citrate pH 5.0 with 150 mM NaCl buffer. Wells were washed extensively and bound LAMP1-RbFc was detected using goat anti-rabbit-IgG(H + L), mouse/human ads-HRP antibody (Southern Biotech, 1:5,500) for 1 h at RT.