Preparation of on-plate immobilized metal ion affinity chromatography platform via dopamine chemistry for highly selective isolation of phosphopeptides with matrix assisted laser desorption/ionization mass spectrometry analysis

• Our method shows high selectivity, sensitivity, and stability in complex samples.
• Our method minimizes the loss of samples.
• There is no need of elution during the process of enrichment.
• Our method realizes high throughput analysis of phosphopeptides enrichment.

In this study, a novel on-plate IMAC technique was developed for highly selective enrichment and isolation of phosphopeptides with high-throughput MALDI-TOF-MS analysis. At first, a MALDI plate was coated with polydopamine (PDA), and then Ti4+ was immobilized on the PDA-coated plate. The obtained IMAC plate was successfully applied to the highly selective enrichment and isolation of phosphopeptides in protein digests and human serum. Because of no loss of samples, the on-plate IMAC platform exhibits excellent selectivity and sensitivity in the selective enrichment and isolation of phosphopeptides, which provides a potential technique for high selectivity in the detection of low-abundance phosphopeptides in biological samples.

In this study, a novel on-plate IMAC technique was developed for highly selective binding of phosphopeptides in protein digests and human serum for high-throughput MALDI-TOF-MS analysis. Thanks to no loss of samples, the on-plate IMAC platform exhibits excellent selectivity and sensitivity in the binding of phosphopeptides. Figure optionsDownload as PowerPoint slide