PhD Scientific Days 2025

Poster Session II. - G: Pharmaceutical Sciences and Health Technologies

A Novel Capillary Electrophoresis Method for Ganglioside Characterization in CNS-Derived Cell Lines

Name of the presenter

Tarnóczi Katinka

Institute/workplace of the presenter

Department of Pharmacodynamics

Authors

Katinka Tarnóczi¹, Éva Szökő¹, Tamás Tábi¹

1: Department of Pharmacodynamics

Text of the abstract

A Novel Capillary Electrophoresis Method for Ganglioside Characterization in CNS-Derived Cell Lines

Katinka Tarnóczi PhD Student; tarnoczi.katinka.timea@semmelweis.hu
Department of Pharmacodynamics, Faculty of Pharmacy, Semmelweis University
Supervisor: Éva Szökő, email: szoko.eva@semmelweis.hu

Introduction: Gangliosides are sialic acid-containing glycosphingolipids involved in key CNS functions such as intercellular signaling, neuronal differentiation, and synapse formation. They contribute to brain development, axonal stability, and myelination. Altered ganglioside expression has been linked to neurodegenerative diseases, neuroinflammation, and tumors. Despite existing analytical techniques, comprehensive ganglioside profiling remains a challenge.
Aims: We aimed to develop a capillary electrophoresis method with laser-induced fluorescence detection (CE-LIF) for the separation of nine biologically relevant, fluorescently labeled gangliosides and apply it to analyze CNS-derived cell lines (C6 glioblastoma, SH-SY5Y neuroblastoma, and BV2 microglia).
Methods: Gangliosides were enzymatically cleaved using endoglycoceramidase I, labeled with APTS, and separated via CE using 15 mM lithium acetate buffer (pH 5.0). Various additives (glycerol, PEO, LPA), pH values, and buffer concentrations were tested to optimize resolution. The method was validated for sensitivity, reproducibility, and accuracy.
Results: Optimal separation was achieved with 5% LPA in 15 mM LiAc buffer, allowing complete separation of nine gangliosides, including the isomers GD1a and GD1b. The method was successfully applied to CNS-derived cell lines. C6 glioblastoma cells predominantly expressed GM3, SH-SY5Y cells showed high levels of GM1, GT1b, and GD1a, while BV2 cells had a unique profile enriched in GM1, GD1a, and GM2.
Conclusion: The validated CE-LIF method enables precise separation and profiling of gangliosides, including isomers. Cell line-specific patterns highlight the potential of gangliosides as biomarkers and their role in CNS pathophysiology.