Phosphopeptide Enrichment for Mass Spectrometry Analysis Using Microscale Titanium Dioxide Solid Phase Extraction

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Jennifer Fournier, Ying Qing Yu, Martin Gilar, Grace M. Credo, Weibin Chen, John C. Gebler
ASMS 2006 May28-June 1 Seattle, Wa
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ASMS 2006, Phosphopeptide Enrichment _TiO2 and IMAC were compared in terms of selectivity and recovery for phosphopeptide enrichment using a standard Enolase peptide/Phosphopeptide mixture.This standard mixture sample allowed us to perform unbiased comparison of different phosphopeptide enrichment devices. The SPE protocol for TiO2 involves less steps than IMAC since metal chelation is not required. For TiO2:The highest recovery from these loading conditions is with the 0.1TFA in high organic.Loading samples mixed with an unique reagent, Enhancer™, ontoTiO2 SPE improves the phosphopeptide selectivity significantly. Additives, such as detergents and salts, at low levels do not alterthe mechanism of action for TiO2, unlike IMAC, and can additionally be used as a sample clean-up device.

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