title: Titanium dioxide nanoparticle coating of polymethacrylate-based chromatographic monoliths for phosphopetides enrichment.
abstract: 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 TiO2nanoparticles onto organic monolithic chromatographic support (CIM-OH-TiO2). We demonstrate the specificity of CIM-OH-TiO2column 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.
authors.affiliation: BIA Separations d.o.o., Mirce 21, SI-5270 Ajdovščina, Slovenia. Electronic address: urh.cernigoj@monoliths.com.
authors.forename: Urh
authors.initials: U
authors.lastname: Černigoj
authors.affiliation: BIA Separations d.o.o., Mirce 21, SI-5270 Ajdovščina, Slovenia. Electronic address: jernej.gaspersic@monoliths.com.
authors.forename: Jernej
authors.initials: J
authors.lastname: Gašperšič
authors.affiliation: Medical University of Vienna, Clinical Department of Laboratory Medicine, Waehringer Guertel 18-20, A-1090 Wien, Austria. Electronic address: andreas.fichtenbaum@meduniwien.ac.at.
authors.forename: Andreas
authors.initials: A
authors.lastname: Fichtenbaum
authors.affiliation: BIA Separations d.o.o., Mirce 21, SI-5270 Ajdovščina, Slovenia. Electronic address: nika.lendero@gmail.com.
authors.forename: Nika
authors.initials: N
authors.lastname: Lendero Krajnc
authors.affiliation: BIA Separations d.o.o., Mirce 21, SI-5270 Ajdovščina, Slovenia. Electronic address: jana.vidic@monoliths.com.
authors.forename: Jana
authors.initials: J
authors.lastname: Vidič
authors.affiliation: Medical University of Vienna, Clinical Department of Laboratory Medicine, Waehringer Guertel 18-20, A-1090 Wien, Austria. Electronic address: goran.mitulovic@meduniwien.ac.at.
authors.forename: Goran
authors.initials: G
authors.lastname: Mitulović
authors.affiliation: BIA Separations d.o.o., Mirce 21, SI-5270 Ajdovščina, Slovenia; The Centre of Excellence for Biosensors, Instrumentation and Process Control - COBIK, Tovarniška 26, SI-5270 Ajdovščina, Slovenia. Electronic address: ales.strancar@monoliths.com.
authors.forename: Aleš
authors.initials: A
authors.lastname: Štrancar
mesh_terms: D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D013058:Mass Spectrometry / Q000379:methods; D008855:Microscopy, Electron, Scanning; D053758:Nanoparticles / Q000737:chemistry*; D010748:Phosphopeptides / Q000032:analysis*; D010766:Phosphorylation; D017550:Spectroscopy, Fourier Transform Infrared; D014025:Titanium / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: D010748:Phosphopeptides; C009495:titanium dioxide; D014025:Titanium
keywords: Chromatography; Metal oxide affinity chromatography; Monolith; Phosphopeptide enrichment; Proteomics; TiO(2) nanoparticles