title: Sample preparation for mass spectrometric analysis of human serum N-glycans using hydrophilic interaction chromatography-based solid phase extraction.
abstract: Expression levels of N-linked glycans derived from human serum glycoproteins have been shown to change during the progression of many diseases. Generally, N-glycans released from human serum proteins co-exist with endogenous serum peptides, salts, and other contaminants. Effective removal of these contaminants is essential to obtain the glycan profile of human serum proteins. Here, we developed a sample preparation method for mass spectrometry (MS) analysis of N-linked glycans derived from human serum glycoproteins based on a zwitterionic hydrophilic material named Click TE-Cys. The high hydrophilicity of Click TE-Cys, resulting from its unique surface structure and charge distribution, facilitated removal of co-existing salts and endogenous serum peptides. Furthermore, the present enrichment approach was handled in parallel, thus saving time. Using this method, a total of 47 unique N-glycans released from human serum proteins were identified. The intrabatch and interbatch coefficients of variation for the 47 N-linked glycans were 8.57% ± 0.96% and 9.22% ± 1.03%, respectively. These results demonstrate that the present method is suitable for fast purification of N-linked glycans derived from human serum glycoproteins, and has potential for clinical application.
authors: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China. yulong@dicp.ac.cn liangxm@dicp.ac.cn.
authors: Liwei
authors: L
authors: Cao
authors: Ye
authors: Y
authors: Zhang
authors: Linlin
authors: L
authors: Chen
authors: Aijin
authors: A
authors: Shen
authors: Xingwang
authors: X
authors: Zhang
authors: Shifang
authors: S
authors: Ren
authors: Jianxin
authors: J
authors: Gu
authors: Long
authors: L
authors: Yu
authors: Xinmiao
authors: X
authors: Liang
mesh_terms: D001798:Blood Proteins / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002853:Chromatography, Liquid / Q000379:methods; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000032:analysis* / Q000097:blood / Q000302:isolation & purification; D052616:Solid Phase Extraction / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D001798:Blood Proteins; D006023:Glycoproteins; D011134:Polysaccharides