Glycosylation of proteins is diverse in structure, widely distributed and exists in various forms such as N-glycosylation, O-GalNAcylation and O-GlcNAcylation. Different glycosylations are involved in a variety of biological processes, including gene transcription, protein translation, cell signaling, cell-cell and host-pathogen interactions. Abnormal changes in glycosylation are related to the occurrence and development of various important diseases, such as immune diseases, cancer, and congenital disorders of glycosylation. This article introduces several common types of glycosylation, the physiological and pathological processes they involved, and the advances in research methods, specifically the enrichment methods, as well as advances in mass spectrometry. The review is expected to broaden the knowledge on glycoproteomics and to provide novel perspectives for the function of glycoproteins, and drug targets or candidate biomarkers discovery.