Abstract: Rapid and effective separation of protein in complex biological samples is very important in the field of life science research, such as biological separation, protein purification and so on. In this study, Fe₃O₄MNP-ZnSQDs were used as carriers, and lysozyme was chosen as template protein. The molecularly imprinted polymer was coated on the surface of the Fe₃O₄MNP-ZnSQDs to form magnetic fluorescent micro-spheres via surface imprinting process. The experiment results show that Fe₃O₄MNP-ZnSQD@MIPs possesses better dispersibility, uniform particle size and strong fluorescence emission, which can be rapidly separated under external magnetic field. Under optimum conditions, the adsorption equilibrium of Fe₃O₄MNP-ZnSQD@MIPs can be achieved in 15 minutes, and the maximum adsorption capacity is 645.76 m g· g^-1, the saturation magnetic intensity is 40 em u· g^-1. Moreover, the prepared Fe₃O₄MNP-ZnSQD@MIPs possesses adsorption selectivity, and the imprinting factor is 2.15. The advantages of the prepared Fe₃O₄MNP-ZnSQD@MIPs include low cost, fast analysis process, convenience and large adsorption capacity. It can be used for rapid separation and purification of lysozyme in complex biological sample.