A novel one-pipeline workflow was reported, which can realize glycoprotein identification and obtain intact glycosylation information after glycopeptide enrichment, without glycan cutting-off.Enriched glycoproteins were digested by Lys-C, and resulted glycopeptides were enriched again and digested by trypsin.Eventually, intact glycopeptides can be utilized for glycosylation analysis, and non-glycopeptides can be used for protein identification.More confident glycosylation information can be obtained from three aspects: improved specificity in glycoprotein identification through two steps of enrichment.