Fig. 2

Optimization of DAS-ELISA reaction conditions (illustrations in color). The maximum positive/negative (P/N) values were selected as the optimal reaction conditions. P/N > 2.1 represents a positive result. A Paired antibody selection. The concentrations of the reagents were as follows: rabbit pAb at 5.67 µg/mL, mAbs at 0.5 µg/mL, and SEE standards at 1 ng/mL. B Optimization of the concentration of pAb as capture antibody. C Optimization of the dilution of mAb as detection antibody. D Optimization of coating conditions. E Blocking agent selection. F Optimization of the reaction time of antigen. G Optimization of the reaction time of detection antibody. H HRP-labeled streptavidin dilution optimization