Poster abstracts
Poster number 32 submitted by Zhefeng Li
Large Scale Purification of Extracellular Vesicle with Reservation of Shape and Function
Zhefeng Li (College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry, Center for RNA Nanobiotechnology and Nanomedicine, Ohio State University), Fengmei Pi (College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry, Center for RNA Nanobiotechnology and Nanomedicine, Ohio State University), Daniel Jasinski (College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry, Center for RNA Nanobiotechnology and Nanomedicine, Ohio State University), Peixuan Guo (College of Pharmacy, College of Medicine, Dorothy M. Davis Heart and Lung Research Institute; NCI Comprehensive Cancer Center; Center for RNA Nanobiotechnology and Nanomedicine, Ohio State University)
Abstract:
Extracellular vesicles (EVs) have been used for non-viral gene delivery therapy. One of the biggest blockage for application is the high cost and low yield of exosomes derived from human cell line. Differential centrifugation is one of the gold standard for EVs purification. However, co-precipitation of large particles can cause impurity of final product and the high centrifuge force in ultracentrifuge step can decompose the EVs on both structure and function. Here we introduce combination of different ultracentrifugation strategy adopted from our previous experience for virus purification to isolate EVs with the reservation of the native structure and function during purification. We compare the size, zeta, electronic microscopic morphology of EVs from HEK293 cell and EVs-like nanovesicles from Ginger. Result indicate EVs purify by the new method has similar majority size but has more homogenous distribution compare to traditional method. TEM image shows cushion method not only remove impurity but also eliminate aggregation and disruption.
Keywords: Extracellular Vesicle, Ultracentrifugation