Preparation of Multi-layer Open Tubular Capillary Columns Coated with Zirconium Phosphonate for Phosphopeptide Enrichment and Identification by Mass Spectrometry

摘要

Satisfactory results for phosphoprotein analysis cannot be easily obtained because of the low abundance of phosphoproteins in cells and the suppressive effect from large amount of non-phosphopeptides during mass spectrometry(MS) analysis. Therefore, open tubular capillary columns (OTCC) coated with one layer of zirconium phosphonate(ZrP) have been demonstrated efficiently in enrichment of phosphopeptides. However, two major disadvantages remain for OTCC: (1) although loading amount can be increased by extending column length, higher back pressure cannot be avoided when handling large amount of sample; (2) phosphopeptide enrichment using OTCC is limited to a narrow flow rate range of mobile phase. To address these issues, poly(amidoamine) multi-layer open tubular capillary columns coated with zirconium phosphonate(ZrP-MOTCC) for phosphopeptide enrichment were developed and demonstrated very well in phosphopeptide enrichment.