Abstract
SummaryA competitive immunoassay for staphylococcal enterotoxin A (SEA) detection in milk was developed, using immobilised antibody onto magnetic nanoparticles (MNPs). MNPs were prepared and then modified to introduce amino groups on them. The morphology and size of the obtained both unmodified and modified MNPs were characterized using TEM analyses. Monoclonal anti‐SEA antibody was immobilised onto the modified MNPs (MNP‐Ab). Staphylococcal enterotoxin A was conjugated with fluorescent dye ATTO620NHS. The characteristics of fluorescence conjugate were examined. The amount of MNP‐Ab and concentration of the fluorescent conjugate used for competitive immunoassay were optimized: 0.25 mg and 53 μg mL−1, respectively. The detection limit of developed immunoassay was determined – 0.23 ng mL−1 SEA in spiked milk samples. The immunoassay takes only 30 min, the magnetic separation is fast (<10 s) and the volume of the sample for analysis is very small (200 μL).
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