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. 2022 Jun 28;11(13):2047. doi: 10.3390/cells11132047

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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The TEA-based HpH-RP fractionation of phosphopeptides. (A) Compared with single-shot LC-MS/MS analysis, HpH-RP fractionation increases the number of phosphopeptides identified from the lactic acid method by nearly two-fold. Three replicates are performed for both single-shot analysis and HpH-RP fractionation. (B) The overlap of the phosphopeptides identified in the three replicates of HPH-RP fractionation. (C) The cumulative number of phosphopeptides and phosphosites identified in 293T cells. (D) The number of phosphopeptides identified and phosphopeptide enrichment specificity in the three fractions of the TEA-based HpH-RP fractionation. Bars show the mean ± SD of three replicates; error bars indicate standard deviation. (E) The separation efficiency of the TEA-based HpH-RP fractionation is shown as the percentage of phosphopeptides found in one or more fractions.