Specific recognition and sensing of glycoproteins are of great importance in clinical diagnostics considering their frequent utilization as biomarkers and therapeutic targets. In this work, a biomimetic fluorescent sensor for the selective and sensitive detection of glycoprotein was developed, which was based on late-model boronate fluorescent molecularly imprinted nanoparticles (B-FMIP NPs). The B-FMIP NPs were fabricated via the macromolecular assembly of a fluorescent photo-crosslinkable amphiphilic copolymer containing boronic acid with glycoprotein in aqueous solution and in situ photo-crosslinking. Due to the synergism of boronate affinity and the molecular imprinting effect, the resultant B-FMIP NPs demonstrated specific recognition and remarkable selectivity toward the template glycoprotein (ovalbumin, OVA) with a high imprinted factor (α) of 6.0 and gave rise to obvious fluorescence quenching after binding with OVA in water. Under optimized experimental conditions, the as-prepared B-FMIP NPs exhibited linearity over the OVA concentration range of 10⁻¹³ to 10⁻³ mg mL⁻¹ with a detection limit of 3.3 × 10⁻¹⁴ mg mL⁻¹, as well as a rapid response time (about 10 min), which was superior to that of other previously reported OVA sensors. Finally, these B-FMIP NPs have been applied for the determination of OVA in real samples.