Site-specific N-glycosylation characterization of micro monoclonal immunoglobulins based on EThcD-sceHCD-MS/MS.
Site-specific N-glycosylation characterization of micro monoclonal immunoglobulins based on EThcD-sceHCD-MS/MS.
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DOI：
10.3389/fimmu.2022.1013990
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发表时间：
2022
影响因子：
7.3
通讯作者：
Zhang, Yong
中科院分区：
文献类型：
--
作者：
Luo, Mengqi;Mao, Yonghong;Zeng, Wenjuan;Zheng, Shanshan;Li, Huixian;Hu, Juanjuan;Xie, Xinfang;Zhang, Yong
由克隆浆细胞产生的单克隆免疫球蛋白是多发性骨髓瘤和具有肾脏意义的单克隆丙种球蛋白病的主要原因。由于纯化方法复杂且纯化的蛋白质和聚糖的化学计量较低，单克隆免疫球蛋白的位点特异性 N-糖基化表征仍然具有挑战性。对单克隆免疫球蛋白的位点特异性 N-糖基化进行分析非常有意义。因此，在本研究中，我们提出了在 HYDRASYS 系统中从多发性骨髓瘤患者中纯化微单克隆 IgA 和 IgG、琼脂糖凝胶内消化、不富集完整 N-糖肽的 LC-MS/MS 分析的集成工作流程，并比较了不同质谱解离方法（EThcD-sceHCD、sceHCD、EThcD 和 sceHCD-pd-ETD）的鉴定性能。结果表明，EThcD-sceHCD 是微量琼脂糖凝胶内免疫球蛋白 (~2 μg) 的位点特异性 N-糖基化表征的更好选择，因为它可以覆盖更多独特的完整 N-糖肽（分别来自 IgA1 和 IgG2 的 37 和 50 个完整 N-糖肽），并提供比 sceHCD、EThcD 和 sceHCD-pd-ETD 更高质量的谱图。我们证明了位点特异性 N-糖基化替代策略的优势，该策略表征了从电泳分离的条带中获得的微单克隆免疫球蛋白。该工作将促进临床N-糖蛋白组学及相关免疫学的发展。
Monoclonal immunoglobulin produced by clonal plasma cells is the main cause in multiple myeloma and monoclonal gammopathy of renal significance. Because of the complicated purification method and the low stoichiometry of purified protein and glycans, site-specific N-glycosylation characterization for monoclonal immunoglobulin is still challenging. To profile the site-specific N-glycosylation of monoclonal immunoglobulins is of great interest. Therefore, in this study, we presented an integrated workflow for micro monoclonal IgA and IgG purification from patients with multiple myeloma in the HYDRASYS system, in-agarose-gel digestion, LC-MS/MS analysis without intact N-glycopeptide enrichment, and compared the identification performance of different mass spectrometry dissociation methods (EThcD-sceHCD, sceHCD, EThcD and sceHCD-pd-ETD). The results showed that EThcD-sceHCD was a better choice for site-specific N-glycosylation characterization of micro in-agarose-gel immunoglobulins (~2 μg) because it can cover more unique intact N-glycopeptides (37 and 50 intact N-glycopeptides from IgA1 and IgG2, respectively) and provide more high-quality spectra than sceHCD, EThcD and sceHCD-pd-ETD. We demonstrated the benefits of the alternative strategy in site-specific N-glycosylation characterizing micro monoclonal immunoglobulins obtained from bands separated by electrophoresis. This work could promote the development of clinical N-glycoproteomics and related immunology.
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DOI：
10.1074/mcp.r120.002204
发表时间：
2021
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Ye Z;Vakhrushev SY
通讯作者：
Vakhrushev SY
通过质谱法进行血清/血浆分析和生物标志物鉴定的样品制备。
DOI：
10.1016/j.chroma.2006.11.054
发表时间：
2007-06-15
期刊：
Journal of chromatography. A
影响因子：
--
作者：
Luque-Garcia JL;Neubert TA
通讯作者：
Neubert TA
影响因子：
4.4
作者：
Riley, Nicholas M.;Malaker, Stacy A.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
影响因子：
4.4
作者：
Sennels, Lau;Salek, Mogjilborahman;Rappsilber, Juri
通讯作者：
Rappsilber, Juri
影响因子：
6.1
作者：
Yu, Guizhen;Zhang, Yong;Zhang, Hong
通讯作者：
Zhang, Hong