Magnetic nanoparticles (MNP) are efficient molecular carriers for affine molecules. MNP complexes with antibodies can be used for the selective concentration and highly sensitive detection of various compounds. In this paper, development steps of the enzyme immunoassay using MNP are considered in details. Simple method of MNP synthesis and production of conjugates of antibodies with the aggregated MNP using physical sorption is presented. Simple, yet effective, formats of enzyme immunoassay are given. Using aflatoxin B1 detection as an example, possibility of decreasing detection limit up to 2 pg/mL, with a considerable decrease in the assay time, and performing immune interaction in the media with high organic content is shown.