Oxidation-Shielded P(St-MMA)@Fe3O4@P(St-MMA) Mesoporous Magnetic Microspheres: A Robust Solid-Phase Carrier for Ultrasensitive CEA Chemiluminescence Immunoassay

Magnetic polymeric microspheres are pivotal solid-phase carriers in chemiluminescence enzyme immunoassays (CLEIA). However, their practical clinical application is frequently hindered by non-specific adsorption, irreversible aggregation, and the intrinsic susceptibility of exposed outermost Fe3O4nanoparticles to oxidation. To overcome these critical bottlenecks, we rationally engineered highly original monodisperseP(St-MMA)@Fe3O4@P(St-MMA) sandwich-structured microspheres. The bespoke amphiphilic outer shell acts as an impenetrable shield against hydration and oxidation, while maintaining a topologically size-matched mesoporous network (average pore size of 13.11 nm) for optimal antibody anchoring. Strikingly, this architecture ensures exceptional long-term colloidal stability, completely preventing macroscopic agglomeration for over six months in buffer solutions. When evaluated in a carcinoembryonic antigen (CEA), CLEIA, our microspheres achieved an ultra-low limit of detection (LOD) of 0.055 ng·mL−1and high analytical recovery (93.37–108.25%). In a head-to-head comparison with industry-standard commercial magnetic beads, the engineered microspheres delivered stronger chemiluminescent signals and lower background noise, demonstrating excellent intra-assay (CV < 4.37%) and inter-assay (CV < 10%) precision. This work establishes a scalable, highly stable materials platform that effectively resolves the persistent oxidation limitations, holding immense practical importance for next-generation ultrasensitive clinical in vitro diagnostics.