Conference Dates
June 12 – 17, 2022
Abstract
Vaccine therapies based on virus-like particles (VLPs) are currently increasing relevance due to the strong immune response they elicit and their manufacture advantages when compared to traditional biopharmaceuticals. During VLP production using mammalian cell-based platforms, different extracellular vesicles (EVs) are coproduced, leading to the need of a complex downstream purification process. Currently there is no effective and efficient method to separate VLPs from EVs which share very similar density and physicochemical properties1,2. Different methods to characterize the EV composition and their protein content and glycosylation signature are used in this work to further understand their biochemical nature. First, a sucrose cushion ultracentrifugation was carried out to isolate the VLP fraction, also containing co-purified EVs. Following, a multiplexed quantitative proteomic approach was used to characterize the VLP-copurified secretome. Three conditions were studied, a non-transfected condition, transiently transfected with an empty plasmid (mock) and with a plasmid for Gag VLP production.
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Recommended Citation
Jesús Lavado-García, Irene González-Domínguez, Laura Cervera, Inmaculada Jorge, Jesús Vázquez, Tao Zhang, Manfred Wuhrer, and Francesc Gòdia, "Differential glycosylation and extracellular vesicle biogenesis in HEK293 upon transient transfection" in "Vaccine Technology VIII", Tarit Mukhopadhyay, Merck Research Laboratories, USA; Charles Lutsch, Sanofi Pasteur, France; Linda Hwee-Lin Lua, University of Queensland, Australia; Francesc Godia, Universitat Autònoma de Barcelona, Spain Eds, ECI Symposium Series, (2022). https://dc.engconfintl.org/vaccine_viii/97