Phosphopeptide enrichment via HPLC? Sign me up! While maybe not the most novel idea in the world, this new paper in press at MCP out of Utrecht describes the feasibility of such an approach. In this analysis, they use an Fe-IMAC column for enrichment. In comparison to tip based approaches they find massively reduced CVs and overall improved reproducibility.
And the yield does not seem to suffer. Using an Orbitrap Velos they find ~9,000 unique phosphopeptides when loading directly onto a single dimension LC separation and were able to push it to ~14,000 when adding SAX and performing 2D separation!
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