Amorphous TiO₂ Nanotubes as a Platform for Highly Selective Phosphopeptide Enrichment

This work reports highly selective phosphopeptide enrichment using amorphous TiO₂ nanotubes (TiO₂NTs) and the same material decorated with superparamagnetic Fe₃O₄ nanoparticles (TiO₂NTs@Fe₃O₄NPs). TiO₂NTs and TiO₂NTs@Fe₃O₄NPs materials were applied for phosphopeptide enrichment both from a simple peptide mixture (tryptic digest of bovine serum albumin and α-casein) and from a complex peptide mixture (tryptic digest of Jurkat T cell lysate). The obtained enrichment efficiency and selectivity for phosphopeptides of TiO₂NTs and TiO₂NTs@Fe₃O₄NPs were increased to 28.7 and 25.3%, respectively, as compared to those of the well-established TiO₂ microspheres. The enrichment protocol was extended for a second elution step facilitating the identification of additional phosphopeptides. It further turned out that both types of amorphous TiO₂ nanotubes provide qualitatively new physicochemical features that are clearly advantageous for highly selective phosphopeptide enrichment. This has been confirmed experimentally resulting in substantial reduction of non-phosphorylated peptides in the enriched samples. In addition, TiO₂NTs@Fe₃O₄NPs combine high selectivity and ease of handling due to the superparamagnetic character of the material. The presented materials and performances are further promising for applications toward a whole range of other types of biomolecules to be treated in a similar fashion.