Fig. 1

Concept of the SAST method. After applying streptavidin to the ELISA plate, biotinylated antigen protein was fixed. The distance between the bottom of the plate and the antigen protein was expected to be at least 10 nm (both arrows), which could maintain the undenatured structure of the protein. Subsequently, the tested antibodies were applied, the unbound antibodies were washed away with PBS, and HRP-conjugated anti-mouse IgG secondary antibodies were added to the wells. After washing the wells, the bound secondary antibodies were visualized as described in the Materials and Methods. This method is expected to allow the mAb screening of antigens with their undenatured structures