Development of hierarchical Fe₃O₄ magnetic microspheres as solid substrates for high sensitive immunoassays

Improving the detection sensitivity of enzyme linked immunosorbent assay (ELISA) is of the utmost importance for meeting the demand of early disease diagnosis. In this work, a sensitive solid substrate for ELISA, i.e., hierarchical iron oxide magnetic microspheres, Fe₃O₄@mSiO₂@poly[poly(ethylene glycol) methacrylate-co-glycidyl methacrylate], was developed via a novel surface-initiated photoiniferter-mediated polymerization (SI-PIMP) strategy. The magnetic microspheres consist of a magnetic Fe₃O₄ core that gives a high magnetic response, a 3D backbone, a mesoporous SiO₂ middle layer, that facilitates microsphere stability and provides anchoring sites, and polymer brushes, that serve as an antifouling and oriented antibody immobilization layer. As a result, the as-prepared microspheres possess a high antibody loading capacity, an enhanced detection signal and a dramatically improved sensitivity, resulting in a 25-fold improvement over conventional ELISA solid substrates.