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Article

Selective TiO2 Phosphopeptide Enrichment of Complex Samples in the Nanogram Range

1
MS Proteomics Research Group, Research Centre for Natural Sciences, 1117 Budapest, Hungary
2
Department of Inorganic and Analytical Chemistry, Budapest University of Technology and Economics, 1111 Budapest, Hungary
3
Hevesy György Ph.D. School of Chemistry, Eötvös Loránd University, 1117 Budapest, Hungary
4
Ph.D. School of Pharmaceutical Sciences, Semmelweis University, 1085 Budapest, Hungary
5
Proteomics Core Facility, Medical University of Vienna, 1090 Vienna, Austria
6
Clinical Department of Laboratory Medicine, Medical University of Vienna, 1090 Vienna, Austria
*
Author to whom correspondence should be addressed.
Shared first authorship.
Separations 2020, 7(4), 74; https://doi.org/10.3390/separations7040074
Received: 17 November 2020 / Revised: 13 December 2020 / Accepted: 15 December 2020 / Published: 17 December 2020
Phosphopeptide enrichment is a commonly used sample preparation step for investigating phosphorylation. TiO2-based enrichment has been demonstrated to have excellent performance both for large amounts of complex and for small amounts of simple samples. However, it has not yet been studied for complex samples in the nanogram range. Our objective was to develop a methodology applicable for complex samples in the low nanogram range, useful for mass spectrometry analysis of tissue microarrays. The selectivity and performance of two stationary phases (TiO2 nanoparticle-coated monolithic column and spin tip filled with TiO2 microspheres) and several loading solvents were studied. Based on this study, we developed an effective and robust method, based on a spin tip with a non-conventional 50 mM citric acid-based loading solvent. It gave excellent results for phosphopeptide enrichment from samples containing a few nanograms of a complex protein mixture. View Full-Text
Keywords: solid-phase extraction; phosphopeptide enrichment; phosphopeptide isolation; phosphorylation; titanium dioxide; metal oxide affinity chromatography solid-phase extraction; phosphopeptide enrichment; phosphopeptide isolation; phosphorylation; titanium dioxide; metal oxide affinity chromatography
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MDPI and ACS Style

Tóth, G.; Bugyi, F.; Sugár, S.; Mitulović, G.; Vékey, K.; Turiák, L.; Drahos, L. Selective TiO2 Phosphopeptide Enrichment of Complex Samples in the Nanogram Range. Separations 2020, 7, 74. https://doi.org/10.3390/separations7040074

AMA Style

Tóth G, Bugyi F, Sugár S, Mitulović G, Vékey K, Turiák L, Drahos L. Selective TiO2 Phosphopeptide Enrichment of Complex Samples in the Nanogram Range. Separations. 2020; 7(4):74. https://doi.org/10.3390/separations7040074

Chicago/Turabian Style

Tóth, Gábor, Fanni Bugyi, Simon Sugár, Goran Mitulović, Károly Vékey, Lilla Turiák, and László Drahos. 2020. "Selective TiO2 Phosphopeptide Enrichment of Complex Samples in the Nanogram Range" Separations 7, no. 4: 74. https://doi.org/10.3390/separations7040074

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