A facile strategy to prepare the core-shell magnetic imprinted nanoparticles for specific recognition of protein is presented. The core is carboxyl-modified Fe3O4 nanoparticles (Fe3O4@COOH) synthesized directly through a simple one-pot hydrothermal method. The formation of imprinted shell with specific recognition cavities includes three steps. Initially, the protein template is non-covalently anchored on Fe3O4@COOH. Next, the chemical oxidation of 3-aminophenylboronic acid takes place on the surface of Fe3O4@COOH-protein complex for further immobilization of template protein. Then, a thin imprinted layer is obtained on Fe3O4@COOH after the removal of the template protein. The resultant imprinted materials not only exhibit good dispersibility, uniform surface morphology, super-paramagnetic property, but also possess fast kinetics, high capacity, as well as favorable selectivity. In addition, the as-synthesized polymers could be used for six cycles of adsorption-desorption without obvious deterioration and applied to specifically separate and enrich target protein from real biological sample successfully. Furthermore, the result of versatility indicates that the facile preparation method is more suitable for basic proteins. (C) 2015 Elsevier B.V. All rights reserved.