Here, Fe3O4 @SiO2 @Poly (DEMA -EGDMA-CO-VPBA) nanoparticles were prepared as a pH-sensitive nanocomposite for adsorption and controlled release of glycoproteins. The average size of particles was found to be 62.5 nm. The ability of these particles to isolate and enrich the glycoprotein Horseradish Peroxidase and nonglycoprotein of Bovine serum was investigated using analytical and spectroscopic methods. Accordingly, the maximum adsorption capacity of nanocomposite for Horseradish Peroxidase was about 129.8 mgg−1 under optimum pH of 9.3. High performance rate, low concentration needed for glycoprotein separation and enrichment and reusable up to five times, are other features of these particles.