Novel bifunctional terbium complex-based nanoparticles were developed using a modified Stber method and a layer-by-layer assembly process. A magnetic core of Fe3O4 nanoparticles was coated with a silica shell to form the first layer. Then a ternary Tb3+ complex (TESPPA-Tb), which acted as a luminescent marker, was covalently bound to the silica surface by stable Si-O-Si bonds. The TESPPA monomer was synthesized by binding pyridine 2,6-dicarboxylic acid to 3-aminopropyltriethoxysilane, which was used as a ligand for coordination with the Tb3+ ions. An outer shell of silica was applied to the nanoparticles to allow for versatility with surface functionalization. The nanoparticles were characterized by X-ray powder diffraction, transmission electron microscopy, Fourier transform infrared spectroscopy, ultravioletvisible spectroscopy, vibration sample magnetometer, and photoluminescence spectroscopy. The bifunctional nanoparticles exhibited favorable superparamagnetic behavior and photoluminescence properties of Tb3+. These nanoparticles have potential applications in biolabeling, bioseparation, immunoassays, and pathogenic diagnosis.