Fig. 3

Optimization of TRFICA reaction conditions (illustrations in color). Values of both positive samples (1 ng/mL SEE) and negative samples (without SEE spiking) were taken into consideration to select the optimal parameters. A Amount of detection antibody (2 µL, 4 µL, 8 µL, 16 µL of 5.67 mg/mL pAb). B Concentration of coating mAbs (1-, 2-, 4-, or eightfold dilution of mAb 1A5). The concentration of mAb 1A5 was 4.13 mg/mL. C Optimization of the sample pad materials (HG-2, SB08, and RB65). D Optimization of the sample volume (80 µL, 100 µL, 120 µL, 150 µL). E Optimization of the detection time (5 min, 8 min, 10 min). T/C is the ratio of the signal intensity of the test line (T line) to the control line (C line)