Department of Biomedical Engineering, Karabük University, Karabük, Türkiye.
Department of Chemistry, Hacettepe University, Ankara, Türkiye.
Biomed Chromatogr. 2022 Dec;36(12):e5503. doi: 10.1002/bmc.5503. Epub 2022 Sep 21.
Glycosylation is a post-translational modification that plays an active role in many cellular events. It also regulates many functions of proteins. Monoclonal antibody-derived drugs are used to treat many diseases, and glycosylation affects the activity of such drugs developed. On the other hand, N-glycans may change in certain diseases. Therefore, rapid and efficient bioanalytical methods are needed for N-glycosylation profiling. The study aimed to develop an integrated stage-tip application for the simple and rapid N-glycosylation profiling of glycoproteins. A fast and inexpensive N-glycosylation profiling was achieved by integrating all glycoproteomic and glycomic sample preparation steps into a stage-tip. The glycomic approach of the integrated stage-tip reduces the N-glycan profiling time from 2 days to approximately 2.5 hours. It also allows the profiling of immunoglobulin G (IgG) N-glycopeptides directly from human plasma. In addition, N-glycosylation profiling can be done in the developed method without sorbents, C or others, such as strong-cation exchange, at the glycopeptide level.
糖基化是一种翻译后修饰，在许多细胞事件中发挥着积极的作用。它还调节蛋白质的许多功能。单克隆抗体衍生的药物被用于治疗许多疾病，糖基化会影响此类药物的活性。另一方面，某些疾病可能会改变 N-糖链。因此，需要快速有效的生物分析方法来进行 N-糖基化分析。本研究旨在开发一种集成的阶段式吸管应用程序，用于简单快速地对糖蛋白进行 N-糖基化分析。通过将所有糖蛋白质组学和糖组学样品制备步骤集成到一个阶段式吸管中，实现了快速且廉价的 N-糖基化分析。集成的阶段式吸管的糖组学方法将 N-糖链分析时间从 2 天缩短至大约 2.5 小时。它还允许直接从人血浆中对免疫球蛋白 G（IgG）N-糖肽进行 N-糖基化分析。此外，在开发的方法中，无需使用吸附剂（如 C 或强阳离子交换剂）即可在糖肽水平上进行 N-糖基化分析。
