A streamlined strategy for rapid and selective analysis of serum N-glycome

Abstract

N-glycome profile of serum has been considered as potential source of biomarker discovery for decades. However, the time-consuming sample preparation and difficulties in N-glycan enrichment limit N-glycome exploration and further applications. Here, a strategy for rapid N-glycome profiling of complex biological samples has been developed. Easy-to-prepare and cost-friendly sterilized cotton is utilized as a matrix for sample absorption, quick enzymatic deglycosylation, and highly selective N-glycan enrichment. Using this multifunctional matrix, a corresponding centrifuge-assisted sample preparation approach and its protocol were developed. Sample preparation could be completed within 2.5 h, while traditional methods usually cost more than 24 h. Additionally, selectivity towards glycan was increased 100 folds than commercially available purification microtips, making the detection more sensitive. Finally, this strategy was successfully applied to analyze the serum N-glycome of colorectal cancer (CRC) patients and healthy individuals, the glycan ratio (Hex5HexNAc4Sia2/Hex5HexNAc4Sia1) was found to significantly increase among CRC patients.