Unlike N-glycans, O-glycans are smaller, highly diversified branched carbohydrate molecules that can attach to various protein structures. This characteristic leads to the complexity of the analysis, making it difficult to analyze O-glycosylation sites. The variability of glycosylation sites is a key indicator of cellular activity, as evidenced by the correlation between the reduction in serum protein glycosylation sites and the severity of congenital disorders of glycosylation (CDGs).