Sun Zhenyu, Fu Bin, Wang Guoli, Zhang Lei, Xu Ruofan, Zhang Ying, Lu Haojie
Shanghai Cancer Center and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China.
Department of Chemistry and NHC Key Laboratory of Glycoconjugates Research, Fudan University, Shanghai 200032, China.
Natl Sci Rev. 2022 Apr 5;10(1):nwac059. doi: 10.1093/nsr/nwac059. eCollection 2023 Jan.
The glycoproteome has emerged as a prominent target for screening biomarkers, as altered glycosylation is a hallmark of cancer cells. In this work, we incorporated tandem mass tag labeling into quantitative glycoproteomics by developing a chemical labeling-assisted complementary dissociation method for the multiplexed analysis of intact -glycopeptides. Benefiting from the complementary nature of two different mass spectrometry dissociation methods for identification and multiplex labeling for quantification of intact -glycopeptides, we conducted the most comprehensive site-specific and subclass-specific -glycosylation profiling of human serum immunoglobulin G (IgG) to date. By analysing the serum of 90 human patients with varying severities of liver diseases, as well as healthy controls, we identified that the combination of IgG1-H3N5F1 and IgG4-H4N3 can be used for distinguishing between different stages of liver diseases. Finally, we used targeted parallel reaction monitoring to successfully validate the expression changes of glycosylation in liver diseases in a different sample cohort that included 45 serum samples.
由于糖基化改变是癌细胞的一个标志，糖蛋白质组已成为筛选生物标志物的一个重要靶点。在这项工作中，我们通过开发一种化学标记辅助的互补解离方法，将串联质量标签标记纳入定量糖蛋白质组学，用于完整糖肽的多重分析。受益于两种不同质谱解离方法用于完整糖肽鉴定的互补性质以及用于定量的多重标记，我们对人血清免疫球蛋白G（IgG）进行了迄今为止最全面的位点特异性和亚类特异性糖基化分析。通过分析90例不同严重程度肝病患者以及健康对照者的血清，我们确定IgG1-H3N5F1和IgG4-H4N3的组合可用于区分肝病的不同阶段。最后，我们使用靶向平行反应监测在一个包含45份血清样本的不同样本队列中成功验证了肝病中糖基化的表达变化。
