Characterization of Extracellular Vesicles Secreted in Lentiviral Producing HEK293SF Cell Cultures

Lentiviral vectors (LVs) are a powerful tool for gene and cell therapy human embryonic kidney cells (HEK293) have been extensively used as platform production of these vectors. Like most cellular tissues, HEK293 release extracellular vesicles (EVs). EVs released by share similar size, biophysical characteristics even biogenesis pathway with cell-produced enveloped viruses, making it challenge to efficiently separate from LVs. Thus, co-purified LVs during downstream processing, considered “impurities” in the context therapy. A greater understanding co-purifying is needed enable improved processing. To that end, an inducible lentivirus producing line were studied under two conditions: non-induced induced. identified both conditions, their presence confirmed transmission electron microscopy Western blot. EV cargos each condition then further characterized multi-omic approach. Nineteen proteins mass spectrometry potential markers differentiate LV preparations. Lipid composition preparations before after induction showed enrichment phosphatidylserine. RNA transcripts involved viral processes binding functions. These findings provide insights on product profile lentiviral could support development separation strategies aimed at removing co-produced EVs.