TiO2 with Confined Water Boosts Ultrahigh Selective Enrichment of Phosphorylated Proteins

In the selective enrichment of phosphorylated proteins (PPs) from biological samples, the non-phosphorylated proteins (NPPs) adhered onto enrichment adsorbents due to the hydrophobic interaction, resulting in poor selectivity and low recovery of target PPs. Herein, superhydrophilic TiO2-coated porous SiO2 microspheres are prepared and boost remarkable selectivity toward standard PP spiked with 2000 mass-fold NPP interference. The outstanding performance of the superhydrophilic microspheres is attributed to the coordination interaction between TiO2 and PPs, and the confined water layer generated from superhydrophilicity avoids the irreversible adsorption of NPPs by keeping NPP inner hydrophobic regions in a compact structure, which is verified by single molecule force spectroscopy, circular dichroism, and quartz crystal microbalance. This strategy for enrichment is expected to solve the challenge in proteomics and sheds light on the interactions between biomolecules and superwettability.