Glycan Node Analysis Detects Varying Glycosaminoglycan Levels in Melanoma-Derived Extracellular Vesicles

Jenifer Pendiuk Goncalves, Sierra A. Walker, Jesús S. Aguilar Díaz de león, Yubo Yang, Irina Davidovich, Sara Busatto, Jann Sarkaria, Yeshayahu Talmon, Chad R. Borges, Joy Wolfram

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


Extracellular vesicles (EVs) play important roles in (patho)physiological processes by mediating cell communication. Although EVs contain glycans and glycosaminoglycans (GAGs), these biomolecules have been overlooked due to technical challenges in comprehensive glycome analysis coupled with EV isolation. Conventional mass spectrometry (MS)-based methods are restricted to the assessment of N-linked glycans. Therefore, methods to comprehensively analyze all glyco-polymer classes on EVs are urgently needed. In this study, tangential flow filtration-based EV isolation was coupled with glycan node analysis (GNA) as an innovative and robust approach to characterize most major glyco-polymer features of EVs. GNA is a molecularly bottom-up gas chromatography-MS technique that provides unique information that is unobtainable with conventional methods. The results indicate that GNA can identify EV-associated glyco-polymers that would remain undetected with conventional MS methods. Specifically, predictions based on GNA identified a GAG (hyaluronan) with varying abundance on EVs from two different melanoma cell lines. Enzyme-linked immunosorbent assays and enzymatic stripping protocols confirmed the differential abundance of EV-associated hyaluronan. These results lay the framework to explore GNA as a tool to assess major glycan classes on EVs, unveiling the EV glycocode and its biological functions.

Original languageEnglish (US)
Article number8506
JournalInternational journal of molecular sciences
Issue number10
StatePublished - May 9 2023


  • extracellular vesicle
  • glycosylation
  • hyaluronic acid
  • mass spectrometry
  • melanoma
  • Humans
  • Hyaluronic Acid/metabolism
  • Glycosaminoglycans/metabolism
  • Melanoma/diagnosis
  • Polysaccharides/metabolism
  • Extracellular Vesicles/metabolism

ASJC Scopus subject areas

  • Molecular Biology
  • Spectroscopy
  • Catalysis
  • Inorganic Chemistry
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry


Dive into the research topics of 'Glycan Node Analysis Detects Varying Glycosaminoglycan Levels in Melanoma-Derived Extracellular Vesicles'. Together they form a unique fingerprint.

Cite this