Titanium dioxide nanoparticle coating of polymethacrylate-based chromatographic monoliths for phosphopetides...

Metal oxide affinity chromatography has been one of the approaches for specific enrichment of phosphopeptides from complex samples, based on specific phosphopeptide adsorption forming bidentate chelates between phosphate anions and the surface of a metal oxide, such as TiO₂, ZrO₂, Fe₂O₃, and Al₂O₃. Due to convective mass transfer, flow-independent resolution and high dynamic binding capacity, monolith chromatographic supports have become important in studies where high resolution and selectivity are required. Here, we report the first synthesis and characterization of immobilisation of rutile TiO₂ nanoparticles onto organic monolithic chromatographic support (CIM-OH-TiO₂). We demonstrate the specificity of CIM-OH-TiO₂ column for enrichment of phosphopeptides by studying chromatographic separation of model phosphorylated and nonphosphorylated peptides as well as proving the phosphopeptide enrichment of digested bovine α-casein. The work described here opens the possibility for a faster, more selective enrichment of phosphopeptides from biological samples that will enable future advances in studying protein phosphorylation.