The copolymer poly(isocyanatoethyl methacrylate-co-methyl methacrylate-co-ethylene glycol dimethacrylate) (poly(IEM-co-MMA-co-EGDMA)) was developed as a novel, facile, highly reactive and versatile monolithic matrix, which was amenable to surface functionalization with a variety of nucleophilic modifiers based on the reactive isocyanate group, producing monoliths of various chemistries suitable for chromatography. The specific surface area, pore size distribution, porosity and morphology of the monolithic matrix were characterized using a mercury intrusion porosimeter and scanning electron microscopy (SEM), respectively. Thermal analysis results revealed that the monolith was thermally stable up to 307 °C. The success of the chemical modification of the monolithic matrix was confirmed by FT-IR, solid state (13) C NMR and XPS elemental analysis, showing the high ligand density of the modified monoliths. A ligand density of up to 2.33 mmol·mL(-1) was obtained for the 1-octanol modified monolith (M1) with an isocyanate group conversion of 96.9%, indicating a high efficiency of the modification reaction. The potential application of the monoliths was demonstrated by the separation of a series of compounds. The novel monolithic columns exhibited high mechanical stability, column efficiency and good repeatability and reproducibility.