PhD Scientific Days 2026

Poster Session 3.H - Pharmaceutical Sciences and Health Technologies

Proteomic and Glycosylation Profiling of Extracellular Vesicles, Cell Membranes, and Cells in Lung Cancer

Name of the presenter

Molnár, Alexandra

Institute/workplace of the presenter

HUN-REN Research Centre for Natural Sciences, MTA-HUN-REN TTK Lendület (Momentum) Glycan Biomarker Research Group

Authors

Alexandra Molnár^1,2, Mirjam Balbisi^1,2, Tamás Langó³, Júlia Németh¹, Beáta G. Vértessy^3,4, Lilla Turiák¹
1: HUN-REN Research Centre for Natural Sciences, MTA-HUN-REN TTK Lendület (Momentum) Glycan Biomarker Research Group
2: Semmelweis University Doctoral School
3: HUN-REN Research Centre for Natural Sciences, Institute of Molecular Life Sciences
4: Budapest University of Technology and Economics, Department of Applied Biotechnology and Food Science, Faculty of Chemical Technology and Biotechnology

Text of the abstract

Lung cancer is one of the most common malignancies worldwide and a leading cause of cancer-related deaths. Its high mortality is largely due to late-stage diagnosis, highlighting the need for sensitive and minimally invasive biomarkers. Extracellular vesicles (EVs), especially small EVs (sEVs), are promising biomarker sources as they reflect the molecular composition of their cells of origin, however, their glycosylation and glycosaminoglycan (GAG) profiles remain poorly understood.

We aimed to perform a comparative analysis of sEVs, cell membranes, and whole cells from three non-small cell lung cancer cell lines (A549: adenocarcinoma, H661: large cell carcinoma, and SKMES-1: squamous cell carcinoma) and a non-cancerous epithelial lung cell line (BEAS-2B).

A549, H661, and SKMES-1 cell lines were cultured in completed RPMI-1640 and BEAS-2B cells were grown in BEGM. For sEV isolation, 24 flasks per cell line were incubated in serum-free medium for 72 hours. The media were collected, bigger particles were removed by centrifugation. and sEVs were isolated using size-exclusion chromatography. For cell and cell membrane preparations, 8 T75 flasks per cell line were used. In the latter case, collected cells were mechanically disrupted, and the cell membranes were pelleted using differential centrifugation.

All four cell lines were successfully cultured and cells, cell membranes, and sEVs were isolated. We observed that BEAS-2B cells showed slower growth than the cancer cell lines and these cells produced a lower yield of sEVs than the cancer cells. Sample preparation workflows, including glycopeptide enrichment and GAG digestion, were systematically optimized prior to analysis. Preliminary results indicate that sEVs are enriched in chondroitin/dermatan sulfate GAGs and glycopeptides compared to whole cells.

This study establishes a platform for comprehensive proteomic, glycoproteomic, and GAG analyses of sEV, whole cell, and cell membrane samples. These investigations are expected to provide deeper knowledge about the molecular characteristics of lung cancer, which could lead to the identification of potential biomarkers and therapeutic targets.

Supported by the Lendület (Momentum) Program of the Hungarian Academy of Sciences.
A. Molnár and M. Balbisi were supported by the Semmelweis 250+ Excellence PhD Scholarship.