Rationally designed peptide conjugated to gold nanoparticles for detection of aflatoxin B1 in point-of-care dot-blot assay

Aflatoxin B1 (AFB1) is a hepatotoxic and carcinogenic food contaminant. Although on-site paper-based detection is sensitive it depends on expensive antibodies which are difficult to raise against mycotoxins. Here, we rationally designed a high binding octapeptide, N-KSGKSKPR-C peptide for AFB1 detection, by molecular docking, as confirmed by indirect ELISA (Kd 323 nM). Further, conjugation of octapeptide with gold nanoparticles (26 nm) permitted its use as a visual detection agent in rapid, sensitive dot-blot assay (LOD 0.39 mu g/kg). The assay displayed negligible cross-reactivity with co-contaminating mycotoxins. AFB1 recovery from spiked wheat sample was comparable by dot-blot (78-91 %) and HPLC (65-87 %). Evaluation of dot-blot using certified reference material and 146 food and feed samples showed high correlation R2 = 0.87 with HPLC. The assay displayed high accuracy (91 %), sensitivity (71 %) and specificity (96.5 %). Therefore, the developed dot-blot assay holds promise for monitoring AFB1 contamination in food and feed.

Automated summary

A high binding octapeptide, N-KSGKSKPR-C, was designed for AFB1 detection, and conjugated with gold nanoparticles to enable visual detection in a rapid dot-blot assay. The developed assay showed high accuracy, sensitivity, and specificity, making it promising for AFB1 monitoring in food and feed.