TY - JOUR
T1 - Exosome-based therapeutics
T2 - Purification using semi-continuous multi-column chromatography
AU - Moleirinho, Mafalda G.
AU - Silva, Ricardo J.S.
AU - Carrondo, Manuel J.T.
AU - Alves, Paula M.
AU - Peixoto, Cristina
PY - 2019/10/1
Y1 - 2019/10/1
N2 - Extracellular vesicles, more particularly exosomes, are nanostructures of great medical interest. Similarly to other complex biopharmaceuticals such as virus, the purification of these nanovesicles is still a considerable challenge. Here we describe a proof-of-concept purification strategy based on the well-established size exclusion chromatography technique, operated in a semi-continuous mode. A two-column system was designed and experimentally validated. Immunoblotting for protein exosome markers and electron microscopy indicated the presence of exosomes, with its characteristic cup-shaped morphology. The isolated nanovesicles have a mode size of 128 nm and a concentration and size distribution very similar between the collected products. Importantly, after comparison with batch chromatography, we were able to increase the yield up to 83%, specific productivities in 167% and a 2.7-fold reduction in buffer consumption with the semi-continuous operation. Finally, we demonstrate that the developed process can be potentially applied in the large-scale production of exosome-based therapeutics.
AB - Extracellular vesicles, more particularly exosomes, are nanostructures of great medical interest. Similarly to other complex biopharmaceuticals such as virus, the purification of these nanovesicles is still a considerable challenge. Here we describe a proof-of-concept purification strategy based on the well-established size exclusion chromatography technique, operated in a semi-continuous mode. A two-column system was designed and experimentally validated. Immunoblotting for protein exosome markers and electron microscopy indicated the presence of exosomes, with its characteristic cup-shaped morphology. The isolated nanovesicles have a mode size of 128 nm and a concentration and size distribution very similar between the collected products. Importantly, after comparison with batch chromatography, we were able to increase the yield up to 83%, specific productivities in 167% and a 2.7-fold reduction in buffer consumption with the semi-continuous operation. Finally, we demonstrate that the developed process can be potentially applied in the large-scale production of exosome-based therapeutics.
KW - Continuous purification
KW - Downstream processing
KW - Exosomes
KW - Extracellular vesicles
KW - Multi-column chromatography
UR - http://www.scopus.com/inward/record.url?scp=85065823449&partnerID=8YFLogxK
U2 - 10.1016/j.seppur.2019.04.060
DO - 10.1016/j.seppur.2019.04.060
M3 - Article
AN - SCOPUS:85065823449
SN - 1383-5866
VL - 224
SP - 515
EP - 523
JO - Separation and Purification Technology
JF - Separation and Purification Technology
ER -