A universal, portable, and ultra-sensitive pipet immunoassay platform for deoxynivalenol detection based on dopamine self-polymerization-mediated bioconjugation and signal amplification

Deoxynivalenol (DON) is highly toxic to the environment and human health. It is important to detect DON with ultra-high sensitivity, ease of operation, and low cost. Inspired by the excellent stability and biocompatibility of polydopamine, a universal, portable and ultra-sensitive pipet immunoassay platform was reported for DON detection based on dopamine self-polymerization (polydopamine coating and polydopamine nanoparticles). The polydopamine coating acted as an effective strategy for biomolecule immobilization on the pipet to improve the coating efficiency that significantly reduced the required concentration of biomolecules. Performing the ELISA in pipets saved nearly 67% of the antigen amount and 83% of the antibody amount, which reduced the detection cost and simplified the experimental steps. The dual signal amplification in this pipet immunoassay enabled ultra-high sensitivity. Polydopamine nanoparticles acted as the enrichment carrier of horseradish peroxidase-goat anti-mouse IgG for the first-round signal amplification, followed by the tyramine-mediated loading of streptavidin-HRP for the second-round signal amplification. The dual-enriched HRP catalyzed the color-developing substrate to achieve highly sensitive colorimetric DON detection with a limit of detection of 0.435 ng/mL.