Cell Reports MethodsMolecular BiologyPeer-reviewed journal article
Exceptional yield vesicle packaged recombinant protein production from E. coli.
Tara A. EastwoodiD, Karen BakeriD, Bree R. StreatheriD, Nyasha AlleniD, Lin WangiD, Stanley W. BotchwayiD, Ian R. BrowniD, Jennifer R. HiscockiD, Christopher LennoniD, Daniel P. MulvihilliD
Abstract
We describe a novel system that exports diverse recombinant proteins in extracellular vesicles from E. coli. The vesicles not only compartmentalise toxic, insoluble and disulphide bond containing proteins in a soluble and functional form (e.g. DNaseI, nanobodies and IgG-fusions), but the continued release of the inducible vesicle packaged proteins into the media supports continuous isolation of protein from active culture within a micro-environment allowing stable long-term storage. This technology results in unprecedented yields of vesicle packaged functional proteins for efficient downstream processing for a wide range of applications from discovery science to applied biotechnology and medicine.
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