Magnetic affinity microspheres with meso-/macroporous shells for selective enrichment and fast separation of phosphorylated biomolecules.

The flowerlike multifunctional affinity microspheres prepared by a facile solvothermal synthesis and subsequent calcination process consist of magnetic cores and hierarchical meso-/macroporous TiO2 shells. The hierarchical porous structure of the flowerlike affinity microspheres is constructed by the macroporous shell from the stacked mesoporous nanopetals which are assembled by small crystallites. The affinity microspheres have a relatively large specific surface area of 50.45 m(2) g(-1) and superparamagnetism with a saturation magnetization (Ms) value of 30.1 emu g(-1). We further demonstrate that they can be applied for rapid and effective purification of phosphoproteins, in virtue of their selective affinity, porous structure, and strong magnetism. In addition, the affinity microspheres can also be used for enrichment of phosphopeptides, and the selectivity is greatly improved due to the increase of mass transport and prevention of the possible "shadow effect" resulting from the smaller and deeper pores by taking advantage of the unique porous structure. Overall, this work will be highly beneficial for future applications in the isolation and identification of phosphorylated biomolecules.