Human serum N‐linked glycans expression levels change during the disease progression. The low abundance, structural diversity, and coexisting matrices hinder their detection in mass spectrometry analysis. Considering the hydrophilic nature of N‐glycans, cellulose/polymer (1,2‐Epoxy‐5‐hexene) nanohybrid is fabricated with oxirane groups functionalized of asparagine to develop solid phase extraction based hydrophilic interaction liquid chromatography sorbent (cellulose/1,2‐Epoxy‐5‐hexene/asparagine). The morphology, elemental analysis, and surface properties are studied through scanning electron microscopy, energy dispersive X‐ray spectroscopy, and Fourier‐transform infrared spectroscopy. The large surface area of cellulose/polymer nanohybrid (2.09 × 102 m2/g) facilitates the high density of asparagine immobilization resulting in better hydrophilic interaction liquid chromatography enrichment under optimized conditions. The enrichment capability of nanohybrid/asparagine is assessed by the N‐Linked glycans released from ovalbumin and immunoglobulin G where 23 and 13 N‐glycans are detected respectively. The nanohybrid/asparagine shows selectivity of 1:1200 with spiked bovine serum albumin and sensitivity down to 100 attomole. Human serum profiling for N‐glycans identifies 52 glycan structures. This new enrichment strategy enriches serum N‐linked glycans in the presence of salts, proteins, endogenous serum peptides, and so forth. [ABSTRACT FROM AUTHOR]