A protocol of carbonized on-column enrichment for urinary exosomal N-glycans profiling

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2023-01, Vol.1215, p.123586-123586, Article 123586
• Main Authors: Chen, Yijie, Deng, Chunhui, Sun, Nianrong
• Format: Article
• Language: English
• Subjects: Capillary trap column; Carbon; Enrichment; Exosome; Glycoproteins - chemistry; Humans; N-glycans; Polysaccharides - chemistry; Urine
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2022.123586

The carbonized packing capillary trap column was proposed for urinary exosomal N-glycan analysis. [Display omitted] •A carbonized packing capillary trap column was prepared for exosome glycan profiling through automatic operation.•Outstanding sensitivity and size-exclusion effect were achieved by the on-column enrichment for both standard and practical samples.•The protocol of carbonized on-column enrichment reveals urinary exosomal N-glycan difference between the normal and patients with bladder carcinoma. As a widely present vesicle, exosome plays an important role in lots of biological processes due to its inclusive cargos. In particular, exosome glycan cargo is attracting attentions since its aberrant alteration is closely related to many progressions in diseases. In this work, a novel carbonized packing capillary trap column for urinary exosomal N-glycan enrichment was proposed. The carbonized packing exhibited large specific surface area, mesoporous structure with narrow pore size distribution and abundant carbon for specially interacting with oligosaccharides. Benefitting from all these advantages, the N-glycans deriving from standard glycoproteins or complex human urine exosomes could be identified with high sensitivity and selectivity. Finally, from the glycans identified in healthy volunteers and patients with bladder carcinoma, we observed that 10 of glycans shared by two groups were obvious downregulation and the 18 were upregulation. These results show great potential of capillary trap column as a tool for the enrichment and detection of glycans in exosomal, attracting more attention on disease progression monitoring and biomarker discovery.