*2.1. Competitive ELISA Conditions*

As a reference method to novel cMID, a cELISA was established using optimized conditions defined by checkerboard titration. Here, aflatoxin B1-BSA conjugate was coated, followed by the application of aflatoxin B1-specific monoclonal antibodies pre-incubated with soluble aflatoxin B1 with concentrations ranging from 0.006 ng·mL−<sup>1</sup> to 5000 ng·mL−<sup>1</sup> sample buffer. Absorbance was measured at 405 nm by indirect readout using a secondary antibody conjugated to horseradish peroxidase (HRPO) targeting mouse antibodies and application of respective substrate.
