Phosphopeptide enrichment and analysis using a combination of metal oxide affinity purification and LC MALDI MS

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Emmanuelle Claude, Marten Snel, Thérèse McKenna, Ying Qing Yu, Charalampos-Philip Iossifidis, John Gebler, and James Langridge
ABRF The Association of Biomolecular Resource Facilities, March 31 - April 3, Tampa, Florida; BSBMB 195th Meeting of the Belgian Society of Biochemistry and Molecular Biology; May 15, 2007, Bruxelles; Hellenics Proteomic Society May 23 - 25, Crete, Greece
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In this study we have evaluated a new way of enriching phosphopeptides by using a metal oxide based solid phase extraction (SPE) micro scale device. This sorbent has a high affinity for phosphopeptides and the problem of acidic adsorption is greatly minimized. Furthermore, recent developments in MALDI and LC-MALDI spotting devices, allow the coupling of the off-line chromatographic separation step to the subsequent MS analysis. This approach has been compared to a homemade microcolumn desalting approach on protein digest mixtures.

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