Abstract
Saliva contains various proteins, particularly abundant are phosphoproteins, that may be related to disease occurrences and that play significant roles in a biological system. Thus, medical diagnostics will benefit tremendously if disease-related protein biomarkers are discovered from saliva. In this paper, we propose and demonstrate an approach using functional zinc oxide coated iron oxide magnetic nanoparticles (Fe3O4@ZnO MNPs) as affinity probes to selectively enrich phosphoproteins from complex saliva samples and as microwave absorbers to assist the enrichment and subsequent tryptic digestion of trapped proteins under microwave heating. The target species trapped by MNPs were characterized by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) combined with protein database search. Entire analysis time was shortened to less than 20 min. The detection limit of this approach for a monophosphopeptide was as low as 250 pM (10 μL).
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Acknowledgements
We thank the National Science Council (NSC) of Taiwan for supporting this work financially. We also thank Tsung-Yi Chen and Ju-Yu Lin for their assistance in preparing the scheme and figures.
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Chen, WY., Chen, YC. Functional Fe3O4@ZnO magnetic nanoparticle-assisted enrichment and enzymatic digestion of phosphoproteins from saliva. Anal Bioanal Chem 398, 2049–2057 (2010). https://doi.org/10.1007/s00216-010-4174-x
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DOI: https://doi.org/10.1007/s00216-010-4174-x