A number of polymer butyl methacrylate-co-ethylene dimethacrylate and lauryl methacrylate-co-ethylene glycol dimethacrylate monolithic phases within capillary columns were produced with integrated gold nano-particle (GNP) modified segments for on-capillary extraction and subsequent separation of biomolecules. Photo-masking and photo-grafting techniques were used to produce an aminated zone at the start of the polymer monolithic columns, which was then converted to a nano-agglomerated section using GNPs, followed by the otherwise unmodified monolith. The extent of amination was verified using scanning capacitively coupled contactless conductivity (sC4D) and the C4D profiles of modified monoliths monitored throughout the entire fabrication process, as a quality control technique. Two approaches to monolith amination were compared for achieving maximum GNP coverage, using surface photo-grafting of either vinyl azlactone or glycidyl methacrylate, prior to reaction with ethylenediamine. The modified monolithic columns were applied to the on-column trapping and subsequent reversed-phase separation of protein standards.