PhD Scientific Days 2024

Poster Session R - Pharmaceutical Sciences and Health Technologies 2.

Proteomic, N-glycomic and Glycosaminoglycan Analysis of A549 and BEAS-2B Cell Line-Derived Small Extracellular Vesicles

Előadó neve

Balbisi, Mirjam, MSc

Neptun code

IG36TI

Előadó munkahelye

HUN-REN Research Centre for Natural Sciences

Előadó telefonszáma

+36702822216

Előadó e-mail címe

balbisi.mirjam@phd.semmelweis.hu

Az előadás címe

Proteomic, N-glycomic and Glycosaminoglycan Analysis of A549 and BEAS-2B Cell Line-Derived Small Extracellular Vesicles

Szerző(k) neve és munkahelye

Mirjam Balbisi^1,2, Tamás Langó³, Virág Horváth¹, Zoltán Varga⁴, Kinga Ilyés⁴, Nikolett Nagy³, Otília Tóth³, Beáta G. Vértessy³, Lilla Turiák¹

1: MTA-TTK Lendület (Momentum) Glycan Biomarker Research Group, HUN-REN Research Centre for Natural Sciences
2: Semmelweis University Doctoral School
3: Institute of Molecular Life Sciences, HUN-REN Research Centre for Natural Sciences
4: Biological Nanochemistry Research Group, HUN-REN Research Centre for Natural Sciences

Bemutatás módja

Poszter

Szekció

Poster Session R - Pharmaceutical Sciences and Health Technologies 2.

Language of the presentation

English

Preferred session

Pharmaceutical Sciences and Health Technologies

Összefoglaló szövege

Introduction: Extracellular vesicles (EVs), including small extracellular vesicles (sEVs), are lipid-bound particles released from cells, playing pivotal roles in cell signaling and intercellular communication. Despite increased research focus on EVs in recent decades, there is a lack of studies addressing the glycosylation of their proteins.
Aim: In our study, we aimed to develop a methodology to examine the proteomic, N-glycopeptide, and chondroitin sulfate glycosaminoglycan profiles of sEVs obtained from A549 lung adenocarcinoma and BEAS-2B non-tumorigenic cell lines.
Method: A549 cells underwent culturing in non-completed F12 medium, while BEAS-2B cells were cultured in BEGM medium for 72 hours. After centrifugation, small EVs were isolated from cell culture supernatant by mini-size exclusion chromatography on in-house prepared columns. Isolated sEVs were characterized by microfluidic resistive pulse sensing and transmission electron microscopy. For nanoUHPLC-MS(MS) analysis, sEVs were subjected to freeze-thaw cycles and digested with trypsin and chondroitinase ABC enzymes to produce peptides and chondroitin sulfate disaccharides, respectively. The peptides obtained were enriched for N-glycopeptides by acetone precipitation.
Results: Proteomic analysis confirmed the presence of about half of the 100 most common vesicular marker proteins, indicating high quality sEV samples. We identified approximately 100 N-glycopeptides per sample, including glycopeptides of vesicular marker proteins, and the most abundant chondroitin sulfate disaccharides.
Conclusion: The developed methodology allows the study of glycosylation of sEV proteins from cancerous and non-cancerous cell lines. Therefore, a study involving 6-6 parallel isolations is currently in preparation to identify biological differences between the two cell lines, which may help to understand the mechanisms underlying disease progression.
Funding: Support of the Lendület (Momentum) Program of the Hungarian Academy of Sciences and Semmelweis 250+ Excellence PhD Scholarship is acknowledged. The Authors are grateful to Markus Ralser for donating the nanoAcquity HPLC to the group.

University

Semmelweis University

Supervisor

Lilla Turiák

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

7468

Start

16:25

End

16:28