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Protein phosphorylation, which is one of the most significant post-translational modification of proteins, plays an important role in many biological processes such as signaling transduction, gene expression and cellular homeostasis.It is of great difficulty in phosphoproteome analysis because of sample complexity and low degree of phosphorylation.In addition, ionization efficiencies and MS signals of phosphopeptides are relatively lower than nonphosphorylated peptides.Therefore, specific enrichment of phosphopeptides before analysis is crucial.