*4.4. Preparation of the ICA Strip*

To prepare the ICA strip for ZEN detection, the various biochemical reagents were immobilized on the corresponding locations. Then, 30 µL of the diluted GAM-IgG or ZEN-antigen were uniformly sprayed on the NC membrane with 5 mm parallel spacing, and were dried for 5 h at 60 ◦C, which were the control line (C line) and test line (T line), respectively. The prepared ZEN-McAb-GNP probes were also uniformly sprayed on the glass-fiber conjugate pad and dried for 1 h at 37 ◦C. The assembly of the

ICA strip for ZEN was the same as the previous technological process. The immobilized sample pad, glass-fiber conjugate pad, NC membrane, and absorbent pad were pasted onto a 30 cm PVC sheet. Then, the assembled pad was cut into a 4 mm wide ICA strip and stored in dry and dark conditions.
