Preparation and Evaluation of Novel Iron Immobilized Metal Affinity Chromatography Monolithic Column

A novel method to prepare Fe3+ immobilized metal affinity chromatography monolithic column was developed, which included in situ polymerization of N-acryloxysuccinimide and ethylene dimethacrylate, nitrilotriacetic acid activation, and Fe3+ immobilization. The monolithic column showed high selectivity, high binding capacity, good recovery, and good reproducibility for the enrichment of phosphopeptides. Such a column was successfully applied for the enrichment of phosphopeptides from the nonfat milk protein digests. All these results show the promising potential of monolithic column for phosphoproteome analysis.