An ionic-liquid-based polymer monolithic column was synthesized by free-radical polymerization within the confines of a stainless-steel column (50 mm × 4.6 mm i.d.). In the processes, ionic liquid and stearyl methacrylate were used as dual monomers, ethylene glycol dimethacrylate as the cross-linking agent, and polyethylene glycol 200 and isopropanol as co-porogens. Effects of the pre-polymerization solution components on the properties of the resulting monoliths were studied in detail. Scanning electron microscopy, nitrogen adsorption-desorption measurements, and mercury intrusion porosimetry were used to investigate the morphology and pore size distribution of the prepared monoliths, which showed that the homemade ionic-liquid-based monolith column possessed a relatively uniform macro-pore structure with a total macropore specific surface area of 44.72 m2  g-1 . Compared to a non-ionic-liquid-based monolith prepared under the same conditions, the ionic-liquid-based monolith exhibited excellent selectivity and high performance for separating proteins from complex bio-samples, such as egg white, snailase, BSA digest solution, human plasma, etc, indicating promising applications in the fractionation and analysis of proteins from the complex bio-samples in proteomics research. This article is protected by copyright. All rights reserved.