Size-Selective Enrichment of N-Linked Glycans Using Highly Ordered Mesoporous Carbon Material and Detection by MALDI-TOF MS

Abstract

Many diseases are characterized by the changes of either glycan structure or glycosylation site of glycoproteins. The glycan profiling can provide the overview of glycosylation in despite of the absence of the glycosylation sites, which in turn simplifies the complexity of disease diagnosis. Herein, we describe a simple method to profile the N-linked glycans by MALDI-TOF MS with the enrichment using oxidized ordered mesoporous carbon, taking advantages of the size-exclusive effect of mesopore against proteins as well as the interaction between glycans and carbon. Twenty four N-linked glycans derived from ovalbumin could be efficiently detected with high signal-to-noise (S/N) ratios and sufficient peak intensities. In the analysis of complex serum samples, 32 N-linked glycans could be profiled, and 5 (4 core-fucosylated glycans) of them were distinguished from liver cancer and healthy samples.

Mesoporous carbon biomaterials

“…Using OMC extraction and 2D LC-MS/MS analysis, Qin et al [95] identified 3402 different endogenous peptides from 20 μL human serum, a much higher number than obtained with MCM-41 mesoporous silica. CMK-3 OMCs yielded high enrichment of N-linked glycans from serum samples due to size-exclusion [96]. In addition, M-HMCNs were designed and synthesized for rapid capture of low-abundance peptides from biosamples based on their high hydrophobicity and rapid magnetic response [97].…”

Recent advances in applications of nanomaterials for sample preparation

“…Besides endogenous peptidome analysis, size-exclusive effect of mesoporous materials was also employed for selective extraction of other LMW species, including glycans [137][138][139] and lipids [80], in the presence of interference HMW proteins. Zou et al [137] prepared oxidized mesoporous carbon for selective enrichment of N-linked glycans after PNGase F cleavage in the presence of residual proteins, and 5 N-linked glycans were identified as potential biomarkers of liver cancer.…”

“…Zou et al [137] prepared oxidized mesoporous carbon for selective enrichment of N-linked glycans after PNGase F cleavage in the presence of residual proteins, and 5 N-linked glycans were identified as potential biomarkers of liver cancer. The oxidized mesoporous carbon was replaced by mesoporous silica-carbon composite nanoparticles NP-MCM-C with mesoporous silica support and carbon modified inner surface to obtain uniform particle shape and better dispersity, and a more sensitive and comprehensive glycan enrichment was accomplished [138].…”

Rapid development of proteomics in China: from the perspective of the Human Liver Proteome Project and technology development

“…Zhang WanJun from Qian XiaoHong's lab [49] developed a highly efficient method for glycan enrichment. Zou HanFa's lab [52] enriched N-linked glycans using OMC. Yang PengYuan's lab [53] developed a method to incorporate 18 O into glycans with endoglycosidase to relatively quantify glycans on modified substrates.…”