Zr(IV)-immobilized Affinity Beads Prepared by Surface Template Polymerization for Capturing Phosphorylated Proteins

Abstract

A novel immobilized metal affinity chromatography (IMAC) bead, Zr(IV)-immobilized resin, was prepared by surface template polymerization to enrich phosphorylated proteins and peptides from complex peptides mixtures. In order to enhance both the kinetics and the efficiency, large pathways for the proteins and peptides in the resin were formed, and the Zr(IV)-phosphate complexes were immobilized on the polymer surface. The morphology of the Zr(IV)-immobilized resin was evaluated the by measuring the specific surface area, pore volume, and pore distribution. The resin possessed large amount of the large-macro pores around 300 nm. The separation performance of β-casein from bovine serum albumin (BSA) solution was evaluated by phosphopeptide enrichment and MALDI-TOF MS analysis. The Zr(IV)-immobilized resin showed the high selectivity of the phosphopeptide.