The fabrication of an ideal noble metal modified magnetic microsphere as high performance SERS substrate that possesses good dispersibility, strong magnetic responsiveness, and high sensitivity is still a challenge. Herein, we reported a novel route to fabricating Ag-coated magnetic core-shell microspheres (Fe 3 O 4 @PEI@Ag) with most of the desired advantages by using polyethyleneimine (PEI) as an interlayer. The size and coverage level of the Ag-NPs shell on Fe 3 O 4 @PEI@Ag microspheres were easily controlled by varying the amount of AgNO 3 .Meanwhile, the magnetic core endowed the Fe 3 O 4 @PEI@Ag microspheres with superior magnetic nature, which enabled convenient separation and further enhanced Raman signals due to enrichment of targeted analytes and abundant interparticle hotspots created by magnetism-induced aggregation. Considering these features, the Fe 3 O 4 @PEI@Ag is expected to be a versatile SERS substrate, which was verified by the detection of adsorbed PATP molecules and human IgG with a detection limit as low as 10 -11 M and 10 -14 g/mL, respectively. Therefore, the novel Fe 3 O 4 @PEI@Ag microsphere has an enormous potential for practical SERS detection applications, especially in the target protein quantitative detection field.Surface-enhanced Raman scattering (SERS) is a powerful fingerprint vibrational spectroscopy with high sensitivity, and it is widely used in biochemical detection, analytical chemistry, environment monitoring and biological sensing. 1-3 All these applications are based on proper utilization of "free-electron-like" metal materials, such as Au, Ag, and Cu, and only these noble metal structures with rough surface can provide obvious SERS effect. 4 Therefore, the key of its prominent application lies in fabricating high performance SERS substrates. So far, researchers have fabricated many SERS substrates, among which metal film over periodic nanostructure and Au/Ag colloids in suspension are extensively utilized. 5 The periodic nanostructure can achieve well sensitivity and high reproducibility, but has disadvantages of high cost and difficult to be further chemically modified. Au/Ag colloidal nanoparticles are more efficient with flexible control over individual geometry, easy to mass fabricate, and have good biocompatibility. 6 However, the drawbacks of Au/Ag colloid substrates are their limited enhancement ability and poor stability. 7 Therefore, a reliable and practical SERS substrate with high sensitivity is still a challenge. It should be noted that, up to now, the main problem of SERS detection is that it is difficult to conduct quantitative analysis and hard to apply in the complex system only depend on the noble metal SERS substrate itself.Magnetic microspheres have been extensively investigated for various applications. 8 Due to their excellent magnetic responsiveness, magnetic microspheres can assist in convenient recycling of novel metals, magnetic enrichment of analytes and rapid separation from reaction system. 9, 10 Recently, noble metal modified magnetic micro...