A Dynamic and Pseudo-Homogeneous MBs-icELISA for the Early Detection of Aflatoxin B₁ in Food and Feed

Aflatoxin B₁ (AFB₁) is one of the most toxic and harmful fungal toxins to humans and animals, and the fundamental way to prevent its entry into humans is to detect its presence in advance. In this paper, the monoclonal antibody mAbA2-2 was obtained via three-step sample amplification and multi-concentration standard detection using a subcloning method based on the limited dilution method with AFB₁ as the target. A dynamic and pseucdo-homogeneous magnetic beads enzyme-linked immunosorbent assay (MBs-icELISA) was established using the prepared antibody as the recognition element and immunomagnetic beads as the antigen carrier. The MBs-icELISA showed good linear correlation in the concentration range of 0.004-10 ng/mL with R² = 0.99396. The limit of detection (LOD) of the MBs-icELISA for AFB₁ was 0.0013 ng/mL. This new ELISA strategy significantly shortened AFB₁ detection time through improved sensitivity compared to the conventional ELISA method.