Abstract:

Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. Therefore, it is very important to establish sensitive methods for high-throughput N-glycan profiling. However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO₂) for the efficient enrichment of protein N-glycans. Firstly, pGMAG-SiO₂ was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. The strong hydrophilicity of the material makes it suitable for the enrichment of N-glycans released from complex samples. Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO₂. Finally, pGMAG-SiO₂ was applied to the enrichment of N-linked glycans from human plasma proteins and 47 glycoforms were successfully identified after enrichment. These results demonstrated the high enrichment efficiency and significant application value of pGMAG-SiO₂ in the analysis of N-glycans.

Key words: N-glycan, N-glycoprotein, core-shell silica packing, surface-initiated atom transfer radical polymerization (SI-ATRP)