Department of Pharmacodynamics, Semmelweis University, 4 Nagyvárad tér, H-1089 Budapest, Hungary.
Molecules. 2024 Aug 9;29(16):3769. doi: 10.3390/molecules29163769.
Gangliosides are sialic acid-containing glycosphingolipids that play an essential role in many biological and pathophysiological processes. They are present in high amounts in the central nervous system and their abnormal metabolism or expression has been observed in many diseases. We have developed and validated a sensitive capillary electrophoresis laser-induced fluorescence (CE-LIF) method for the separation and quantification of oligosaccharides digested from nine gangliosides of high biological relevance. APTS was used for the labeling of the glycans. Reverse polarity CE was performed for the separation of the labeled glycans bearing negative charges. The optimized background electrolyte is a 15 mM lithium acetate buffer with pH of 5 containing 5% / linear polyacrylamide, which allows for the separation of all nine gangliosides. Validation parameters including linearity, precision, and accuracy were evaluated. LOQ and LOD were in the nM range, comparable to those of LC-MS techniques. The method was used to identify and quantify the ganglioside pattern of glioblastoma and neuroblastoma cell lines. The presented method is a valuable tool for further investigations aiming at understanding the role of gangliosides in various neurological diseases or CNS tumors.
神经节苷脂是含有唾液酸的糖鞘脂，在许多生物学和病理生理学过程中发挥着重要作用。它们在中枢神经系统中含量丰富，在许多疾病中观察到其代谢或表达异常。我们开发并验证了一种灵敏的毛细管电泳激光诱导荧光（CE-LIF）方法，用于分离和定量从具有高生物学相关性的九种神经节苷脂中消化得到的寡糖。APTS 用于聚糖的标记。采用反相 CE 分离带负电荷的标记聚糖。优化的背景电解质是 15mM 乙酸锂缓冲液，pH 值为 5，其中含有 5%/线性聚丙烯酰胺，可分离所有九种神经节苷脂。评估了线性度、精密度和准确度等验证参数。LOQ 和 LOD 在纳摩尔范围内，与 LC-MS 技术相当。该方法用于鉴定和定量神经胶质瘤和神经母细胞瘤细胞系的神经节苷脂模式。所提出的方法是进一步研究旨在了解神经节苷脂在各种神经疾病或中枢神经系统肿瘤中的作用的有价值的工具。
