A full-automated magnetic particle-based chemiluminescence immunoassay for rapid detection of cortisol in milk

A sensitive full-automated micromagnetic particles (MMPs) based competitive chemiluminescent immunoassay (CLIA) was developed to detect cortisol in milk. Polyclonal antibody (pAb) with good specificity against cortisol was produced. The antigen (cortisol-OVA) was labeled with acridinium ester (cortisol-OVA-AE) as signal tracer. During the detection, the free cortisol in sample will compete with cortisol-OVA-AE for binding to pAb. To capture pAb, MMPs conjugated with goat anti-rabbit IgG was added. The whole immunoassay process (exclude sample pretreatment) was performed by automatic chemiluminescence immunoassay instrument, which could consume less test time (within 40 min) and avoid error from manual operation. The method showed a good detection limit of 0.12 ng/mL, a broad linear range from 0.42 to 72.27 ng/mL for cortisol detection, negligible cross-reactivity with related analogues and satisfied recovery (84.3%–102.3%) for spiked milk samples test. Simultaneously, since the results of proposed method had no significant difference with those of LC–MS/MS, the proposed method was confirmed to have a potential applicability for rapidly monitoring cortisol in the food. [Display omitted] •An automatic rapid assay for cortisol in milk was developed using MMPs-based CLIA.•Cortisol detection: 0.42–72.27 ng/mL concentration range; 0.12 ng/mL detection limit.•The proposed method showed high sensitivity, good reproducibility and stability.•The strategy showed great potential in the fabrication of a novel cortisol test kits.