Magnetic nanoagent assisted deciphering of heterogeneous glycans in extracellular vesicles of varied cellular origins

Extracellular vesicles bear a rich glycome that presents versatile functions in diverse biological processes. Leverage polydopamine modified magnetic particles to serve as nanosized agents for rapid and robust EV capture and manipulation, we here integrated the easy magnetic actuation with specific lectin-glycan binding and enzyme-mediated fluorescence amplification and thus proposed a facile approach to efficiently decipher a broad spectrum of glycans in EVs. Termed magnetic nanoagent assisted extracellular vesicle glycan deciphering (MAEG), the developed assay utilized a magnet as the assistant operation tool and realized fast (∼1 h) and sensitive (a limit of detection of ∼0.7 μg/mL vesicles) EV glycan analysis in a simple low-cost (around 2.27 Chinese Yuan for one test) manner without requirement of any sophisticated platforms. With robust performance for different sample species, the assay achieved to depict the comprehensive glycosylation landscapes for varied EVs derived from eight cell lines focusing on non-small-cell lung cancer. Systematic analyses clearly revealed the high heterogeneity in glycan features of EVs of varied cellular origins. Using an established difference network method, unique glycan features in different EVs were sifted out and further compiled to construct lectin-denoted patterns as dedicated glycosylation fingerprints, potentially expanding EV-based clinical applications.