*4.7. cMID Calibration Curve Analysis*

For cMID calibration curve experiments, a pre-incubation of free mycotoxin and biotinylated antibody was performed. For this, serially diluted aflatoxin B1 samples in PBS, with concentrations ranging from 0.006 ng·mL−<sup>1</sup> to 100,000 ng·mL−<sup>1</sup> , were mixed 1:1 with 225 µL biotinylated antibody, also diluted in PBS. After incubation of one hour at room temperature in a dark surrounding, 500 µL of each sample was applied on coated and blocked columns and also incubated as mentioned above. After washing each column twice with 750 <sup>µ</sup>L PBS, 80 <sup>µ</sup>g·mL−<sup>1</sup> of 70 nm or 700 nm magnetic particles were applied and incubated, as mentioned before. After washing twice, the readout was done as described above.
